7-deazapurine nucleosides for therapeutic uses

ABSTRACT

The invention provides compounds of formula I, wherein R1, R2 and R3 have values defined in the specification and a pharmaceutically acceptable salt thereof; or an optical isomer thereof; or a mixture of optical isomers, as well as compositions comprising such compounds and therapeutic methods that utilize such compounds and/or compositions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 U.S.C. §371 of PCTApplication No. PCT/CZ2010/00050, filed Apr. 19, 2010, which claimspriority to U.S. Provisional Application No. 61/171,656, filed Apr. 22,2009, each of which is incorporated in its entirety herein for allpurposes.

BACKGROUND OF THE INVENTION

The present invention relates to novel anti-proliferation compounds andtheir therapeutic uses.

SUMMARY OF THE INVENTION

The present invention provides anti-cancer compounds. Accordingly, inone aspect the present invention provides a compound of the invention,which is a compound of formula I:

wherein:

R₁ is hydrogen, mono-, di-, or tri-phosphate;

R₂ is aryl, heteroaryl, or alkynyl, wherein said aryl is optionallysubstituted by one or two substituents selected from the groupconsisting of alkoxy, alkylthio, or halogen;

R₃ is hydrogen or alkyl; or

a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ is arylthat is optionally substituted by one substituent selected from thegroup consisting of alkoxy, alkylthio, or halogen; R₃ is hydrogen; or apharmaceutically acceptable salt thereof; or an optical isomer thereof;or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ is arylthat is optionally substituted by one substituent selected from thegroup consisting of alkoxy, alkylthio, or halogen; R₃ is alkyl; or apharmaceutically acceptable salt thereof; or an optical isomer thereof;or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ isheteroaryl; R₃ is hydrogen, with the proviso that R₂ is not1,3-oxazol-2-yl, furan-2-yl, 1,2,4-triazin-3-yl,5,6-dimethyl-1,2,4-triazin-3-yl, 5,6-diphenyl-1,2,4-triazin-3-yl,1,2,4-oxadiazol-3-yl, 4H-1,2,4-triazol-3-yl,5-thioxo-4,5-dihydro-1H-1,2,4-triazol-3-yl,4,5-dihydro-1H-imidazol-2-yl, 4-phenylthiazol-2-yl, 1H-tetrazol-5-yl,1,4,5,6-tetrahydropyrimidin-2-yl, or9-oxo-9H-indeno[1,2-e][1,2,4]triazin-3-yl;

or a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ isheteroaryl; R₃ is alkyl, with the proviso that R₂ is not1,3-oxazol-2-yl, furan-2-yl, 1,2,4-triazin-3-yl,5,6-dimethyl-1,2,4-triazin-3-yl, 5,6-diphenyl-1,2,4-triazin-3-yl,1,2,4-oxadiazol-3-yl, 4H-1,2,4-triazol-3-yl,5-thioxo-4,5-dihydro-1H-1,2,4-triazol-3-yl,4,5-dihydro-1H-imidazol-2-yl, 4-phenylthiazol-2-yl, 1H-tetrazol-5-yl,1,4,5,6-tetrahydropyrimidin-2-yl, or9-oxo-9H-indeno[1,2-e][1,2,4]triazin-3-yl;

or a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ isalkynyl; R₃ is hydrogen; or a pharmaceutically acceptable salt thereof;or an optical isomer thereof; or a mixture of optical isomers.

The invention also provides a pharmaceutical composition comprising acompound of formula I, or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient.

The invention also provides a method of inhibiting tumor growth or cellproliferation in tumor/cancer cells in vitro or in vivo comprisingcontacting a subject in need of such treatment with a compound offormula I, or a pharmaceutically acceptable salt thereof.

The invention also provides a method of treating cancer in an animalcomprising administering to said animal a compound of formula I, or apharmaceutically acceptable salt thereof.

The invention also provides a method of inhibiting a neoplastic diseasein an animal comprising, administering to said animal a compound offormula I, or a pharmaceutically acceptable salt thereof.

The invention also provides the use of a compound of formula I, or apharmaceutically acceptable salt thereof, to prepare a medicament forinhibiting tumor/cancer cell growth or cell proliferation intumor/cancer cells, slowing down cell cycle progression in tumor/cancercells, and for treating cancer in an animal.

The invention also provides the use of a compound of formula I, or apharmaceutically acceptable salt thereof, to prepare a medicament forinhibiting a neoplastic disease in an animal.

The invention also provides synthetic processes and syntheticintermediated disclosed herein that are useful for preparing compoundsof formula (I) or salts thereof

DETAILED DESCRIPTION

For purposes of interpreting this specification, the followingdefinitions will apply and whenever appropriate, terms used in thesingular will also include the plural and vice versa.

As used herein, the term “alkyl” refers to a branched or unbranchedhydrocarbon moiety. Preferably the alkyl comprises 1 to 20 carbon atoms,more preferably 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 7carbon atoms, or 1 to 4 carbon atoms. Representative examples of alkylinclude, but are not limited to, methyl, ethyl, n-propyl, iso-propyl,n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl,neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl,2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl and the like.When an alkyl group includes one or more unsaturated bonds, it may bereferred to as an alkenyl (double bond) or an alkynyl (triple bond)group. Furthermore, when an alkyl group is linked to an aryl group(defined below), it may be referred to as an “arylalkyl” group.

As used herein, the term “alkynyl” refers to both straight- andbranched-chain hydrocarbon groups of 2 to 12 carbon atoms having one ormore carbon-carbon triple bonds. Preferably the alkynyl contains 2 to 8or 2 to 4 carbon atoms. Non-limiting examples of alkynyl includesethynyl, propynyl, butynyl, octynyl, decynyl, and the like.

As used herein, the term “alkoxy” refers to alkyl-O—, wherein alkyl isdefined herein above. Representative examples of alkoxy include, but arenot limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy,tert-butoxy, pentyloxy, hexyloxy, cyclopropyloxy-, cyclohexyloxy- andthe like. As used herein, the term “lower alkoxy” refers to the alkoxygroups having 1-7 carbons and preferably 1-4 carbons.

The term “aryl” refers to monocyclic or bicyclic aromatic hydrocarbongroups having 6-20 carbon atoms in the ring portion. Preferably, thearyl is a (C₆-C₁₀) aryl. Non-limiting examples include phenyl, biphenyl,naphthyl or tetrahydronaphthyl, each of which may optionally besubstituted by 1-4 substituents, such as optionally substituted alkyl,trifluoromethyl, cycloalkyl, halo, hydroxy, alkoxy, acyl, alkyl-C(O)—O—,aryl-O—, heteroaryl-O—, optionally substituted amino, thiol, alkylthio,arylthio, nitro, cyano, carboxy, alkyl-O—C(O)—, carbamoyl, alkylthiono,sulfonyl, sulfonamido, heterocycloalkyl and the like.

Furthermore, the term “aryl” as used herein, also refers to an aromaticsubstituent which can be a single aromatic ring, or multiple aromaticrings that are fused together, linked covalently, or linked to a commongroup such as a methylene or ethylene moiety. The common linking groupalso can be a carbonyl as in benzophenone or oxygen as in diphenyletheror nitrogen as in diphenylamine.

As used herein, the term “heteroaryl” refers to a 5-14 memberedmonocyclic- or bicyclic- or fused polycyclic-ring system, having 1 to 8heteroatoms selected from N, O, S or Se. Preferably, the heteroaryl is a5-10 membered ring system. Typical heteroaryl groups include 2- or3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl, 2-, 4-, or 5-imidazolyl, 3-,4-, or 5-pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-, 4-, or 5-isothiazolyl,2-, 4-, or 5-oxazolyl, 3-, 4-, or 5-isoxazolyl, 3- or 5-1,2,4-triazolyl,4- or 5-1,2,3-triazolyl, tetrazolyl, 2-, 3-, or 4-pyridyl, 3- or4-pyridazinyl, 3-, 4-, or 5-pyrazinyl, 2-pyrazinyl, 2-, 4-, or5-pyrimidinyl.

The term “heteroaryl” also refers to a group in which a heteroaromaticring is fused to one or more aryl, cycloaliphatic, or heterocycloalkylrings, where the radical or point of attachment is on the heteroaromaticring. Nonlimiting examples include but are not limited to 1-, 2-, 3-,5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-,3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-,4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinoliyl, 1-, 3-, 4-, 5-,6-, 7-, or 8-isoquinoliyl, 1-, 4-, 5-, 6-, 7-, or 8-phthalazinyl, 2-,3-, 4-, 5-, or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7-, or 8-quinazolinyl,3-, 4-, 5-, 6-, 7-, or 8-cinnolinyl, 2-, 4-, 6-, or 7-pteridinyl, 1-,2-, 3-, 4-, 5-, 6-, 7-, or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-,or 8-carbzaolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-carbolinyl, 1-, 2-,3-, 4-, 6-, 7-, 8-, 9-, or 10-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-,7-, 8-, or 9-acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-perimidinyl,2-, 3-, 4-, 5-, 6-, 8-, 9-, or 10-phenathrolinyl, 1-, 2-, 3-, 4-, 6-,7-, 8-, or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenoxazinyl,2-, 3-, 4-, 5-, 6-, or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, or10-benzisoqinolinyl, 2-, 3-, 4-, or thieno[2,3-b]furanyl, 2-, 3-, 5-,6-, 7-, 8-, 9-, 10-, or 11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6-,or 7-2H-furo[3,2-b]-pyranyl, 2-, 3-, 4-, 5-, 7-, or8-5H-pyrido[2,3-d]-o-oxazinyl, 1-, 3-, or 5-1H-pyrazolo[4,3-d]-oxazolyl,2-, 4-, or 54H-imidazo[4,5-d]thiazolyl, 3-, 5-, or8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5-, or 6-imidazo[2,1-b]thiazolyl,1-, 3-, 6-, 7-, 8-, or 9-furo[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-,8-, 9-, 10, or 11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6-, or7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-, 4-, 5-, 6-,or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-benzoxapinyl, 2-,4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-,or 11-1H-pyrrolo[1,2-b][2]benzazapinyl. Typical fused heteroaryl groupsinclude, but are not limited to 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolinyl,1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-, 5-, 6-, or7-benzothiazolyl.

A heteroaryl group may be mono-, bi-, tri-, or polycyclic, preferablymono-, bi-, or tricyclic, more preferably mono- or bicyclic.

As used herein, the term “halo” or “halogen” refers to fluoro, chloro,bromo, and iodo.

As used herein, the term “isomers” refers to different compounds thathave the same molecular formula. Also as used herein, the term “anoptical isomer” refers to any of the various stereo isomericconfigurations which may exist for a given compound of the presentinvention and includes geometric isomers. It is understood that asubstituent may be attached at a chiral center of a carbon atom.Therefore, the invention includes enantiomers, diastereomers orracemates of the compound. “Enantiomers” are a pair of stereoisomersthat are non-superimposable mirror images of each other. A 1:1 mixtureof a pair of enantiomers is a “racemic” mixture. The term is used todesignate a racemic mixture where appropriate. “Diastereoisomers” arestereoisomers that have at least two asymmetric atoms, but which are notmirror-images of each other. The absolute stereochemistry is specifiedaccording to the Cahn-Ingold-Prelog R-S system. When a compound is apure enantiomer the stereochemistry at each chiral carbon may bespecified by either R or S. Resolved compounds whose absoluteconfiguration is unknown can be designated (+) or (−) depending on thedirection (dextro- or levorotatory) which they rotate plane polarizedlight at the wavelength of the sodium D line. Certain of the compoundsdescribed herein contain one or more asymmetric centers and may thusgive rise to enantiomers, diastereomers, and other stereoisomeric formsthat may be defined, in terms of absolute stereochemistry, as (R)- or(S). The present invention is meant to include all such possibleisomers, including racemic mixtures, optically pure forms andintermediate mixtures. Optically active (R)- and (S)-isomers may beprepared using chiral synthons or chiral reagents, or resolved usingconventional techniques. If the compound contains a double bond, thesubstituent may be E or Z configuration. If the compound contains adisubstituted cycloalkyl, the cycloalkyl substituent may have a cis- ortrans-configuration. All tautomeric forms are also intended to beincluded.

As used herein, the term “pharmaceutically acceptable salts” refers tosalts that retain the biological effectiveness and properties of thecompounds of this invention and, which are not biologically or otherwiseundesirable. In many cases, the compounds of the present invention arecapable of forming acid and/or base salts by virtue of the presence ofamino and/or carboxyl groups or groups similar thereto (e.g., phenol orhydroxyamic acid). Pharmaceutically acceptable acid addition salts canbe formed with inorganic acids and organic acids. Inorganic acids fromwhich salts can be derived include, for example, hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and thelike. Organic acids from which salts can be derived include, forexample, acetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like. Pharmaceutically acceptable base additionsalts can be formed with inorganic and organic bases. Inorganic basesfrom which salts can be derived include, for example, sodium, potassium,lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese,aluminum, and the like; particularly preferred are the ammonium,potassium, sodium, calcium and magnesium salts. Organic bases from whichsalts can be derived include, for example, primary, secondary, andtertiary amines, substituted amines including naturally occurringsubstituted amines, cyclic amines, basic ion exchange resins, and thelike, specifically such as isopropylamine, trimethylamine, diethylamine,triethylamine, tripropylamine, and ethanolamine. The pharmaceuticallyacceptable salts of the present invention can be synthesized from aparent compound, a basic or acidic moiety, by conventional chemicalmethods. Generally, such salts can be prepared by reacting free acidforms of these compounds with a stoichiometric amount of the appropriatebase (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, or thelike), or by reacting free base forms of these compounds with astoichiometric amount of the appropriate acid. Such reactions aretypically carried out in water or in an organic solvent, or in a mixtureof the two. Generally, non-aqueous media like ether, ethyl acetate,ethanol, isopropanol, or acetonitrile are preferred, where practicable.Lists of additional suitable salts can be found, e.g., in Remington'sPharmaceutical Sciences, 20th ed., Mack Publishing Company, Easton, Pa.,(1985), which is herein incorporated by reference.

As used herein, the term “pharmaceutically acceptable carrier/excipient”includes any and all solvents, dispersion media, coatings, surfactants,antioxidants, preservatives (e.g., antibacterial agents, antifungalagents), isotonic agents, absorption delaying agents, salts, drugs, drugstabilizers, binders, excipients, disintegration agents, lubricants,sweetening agents, flavoring agents, dyes, such like materials andcombinations thereof, as would be known to one of ordinary skill in theart (see, for example, Remington's Pharmaceutical Sciences, 18th Ed.Mack Printing Company, 1990, pp. 1289-1329, incorporated herein byreference). Except in so far as any conventional carrier is incompatiblewith the active ingredient, its use in the therapeutic or pharmaceuticalcompositions is contemplated.

The term “therapeutically effective amount” of a compound of the presentinvention refers to an amount of the compound of the present inventionthat will elicit the biological or medical response of a subject, orameliorate symptoms, slow or delay disease progression, or prevent adisease, etc. In a preferred embodiment, the “effective amount” refersto the amount that inhibits or reduces proliferation of cancer cells, orinhibiting or reducing tumor/cancer growth in vitro or in vivo, orinhibiting or reducing a neoplastic disease in a subject such as amammal. In another preferred embodiment, it also refers to the amountthat reduces the primary tumor/cancer size, inhibits cancer cellinfiltration into peripheral organs, slows or stops tumor metastasis, orrelieves at least to some extent one or more symptoms associated withtumor or cancer, etc. . . .

As used herein, the term “subject” refers to an animal. Preferably, theanimal is a mammal. A subject also refers to for example, primates(e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats,mice, fish, birds and the like. In a preferred embodiment, the subjectis a human.

As used herein, the term “a disorder” or “a disease” refers to anyderangement or abnormality of function; a morbid physical or mentalstate. See Dorland's Illustrated Medical Dictionary, (W.B. Saunders Co.27th ed. 1988).

As used herein, the term “inhibition” or “inhibiting” refers to thereduction or suppression of a given condition, symptom, or disease, or asignificant decrease in the baseline activity of a biological activityor process. In one embodiment, it refers to ability to cause reductionof a tumor or cancer growth, or reduction of the tumor or cancer size.

As used herein, the term “treating” or “treatment” of any disease ordisorder refers in one embodiment, to ameliorating the disease ordisorder (i.e., arresting or reducing the development of the disease orat least one of the clinical symptoms thereof). In another embodiment“treating” or “treatment” refers to ameliorating at least one physicalparameter, which may not be discernible by the patient. In yet anotherembodiment, “treating” or “treatment” refers to modulating the diseaseor disorder, either physically, (e.g., stabilization of a discerniblesymptom), physiologically, (e.g., stabilization of a physicalparameter), or both. In yet another embodiment, “treating” or“treatment” refers to preventing or delaying the onset or development orprogression of the disease or disorder.

As used herein, the term “a,” “an,” “the” and similar terms used in thecontext of the present invention (especially in the context of theclaims) are to be construed to cover both the singular and plural unlessotherwise indicated herein or clearly contradicted by the context.Recitation of ranges of values herein is merely intended to serve as ashorthand method of referring individually to each separate valuefalling within the range. Unless otherwise indicated herein, eachindividual value is incorporated into the specification as if it wereindividually recited herein. All methods described herein can beperformed in any suitable order unless otherwise indicated herein orotherwise clearly contradicted by context. The use of any and allexamples, or exemplary language (e.g. “such as”) provided herein isintended merely to better illuminate the invention and does not pose alimitation on the scope of the invention otherwise claimed. No languagein the specification should be construed as indicating any non-claimedelement essential to the practice of the invention.

The present invention provides anti-cancer compounds. Accordingly, inone aspect the present invention provides a compound of the invention,which is a compound of formula I:

wherein:

R₁ is hydrogen, mono-, di-, or tri-phosphate;

R₂ is aryl, heteroaryl, or alkynyl, wherein said aryl is optionallysubstituted by one or two substituents selected from the groupconsisting of alkoxy, alkylthio, or halogen;

R₃ is hydrogen or alkyl; or

a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ is arylthat is optionally substituted by one substituent selected from thegroup consisting of alkoxy, alkylthio, or halogen; R₃ is hydrogen; or apharmaceutically acceptable salt thereof; or an optical isomer thereof;or a mixture of optical isomers.

Preferably, the present invention provides a compound of formula (I), inwhich R₁ is hydrogen, R₂ is phenyl that is optionally substituted by onesubstituent selected from the group consisting of (C₁-C₄) alkoxy,(C₁-C₄) alkylthio, or halogen; R₃ is hydrogen; or a pharmaceuticallyacceptable salt thereof; or an optical isomer thereof; or a mixture ofoptical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ is arylthat is optionally substituted by one substituent selected from thegroup consisting of alkoxy, alkylthio, or halogen; R₃ is alkyl; or apharmaceutically acceptable salt thereof; or an optical isomer thereof;or a mixture of optical isomers.

Preferably, the present invention provides a compound of formula (I), inwhich R₁ is hydrogen, R₂ is phenyl that is optionally substituted by onesubstituent selected from the group consisting of (C₁-C₄) alkoxy,(C₁-C₄) alkylthio, or halogen; R₃ is (C₁-C₄) alkyl; or apharmaceutically acceptable salt thereof; or an optical isomer thereof;or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ isheteroaryl; R₃ is hydrogen, with the proviso that R₂ is not1,3-oxazol-2-yl, furan-2-yl, 1,2,4-triazin-3-yl,5,6-dimethyl-1,2,4-triazin-3-yl, 5,6-diphenyl-1,2,4-triazin-3-yl,1,2,4-oxadiazol-3-yl, 4H-1,2,4-triazol-3-yl,5-thioxo-4,5-dihydro-1H-1,2,4-triazol-3-yl,4,5-dihydro-1H-imidazol-2-yl, 4-phenylthiazol-2-yl, 1H-tetrazol-5-yl,1,4,5,6-tetrahydropyrimidin-2-yl, or9-oxo-9H-indeno[1,2-e][1,2,4]triazin-3-yl;

or a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

Preferably, the present invention provides a compound of formula (I), inwhich R₁ is hydrogen, R₂ is (5-7) membered heteroaryl; R₃ is hydrogen,with the proviso that R₂ is not 1,3-oxazol-2-yl, furan-2-yl,1,2,4-triazin-3-yl, 5,6-dimethyl-1,2,4-triazin-3-yl,5,6-diphenyl-1,2,4-triazin-3-yl, 1,2,4-oxadiazol-3-yl,4H-1,2,4-triazol-3-yl, 5-thioxo-4,5-dihydro-1H-1,2,4-triazol-3-yl,4,5-dihydro-1H-imidazol-2-yl, 4-phenylthiazol-2-yl, 1H-tetrazol-5-yl,1,4,5,6-tetrahydropyrimidin-2-yl, or9-oxo-9H-indeno[1,2-e][1,2,4]triazin-3-yl;

or a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ isheteroaryl; R₃ is alkyl, with the proviso that R₂ is 1,3-oxazol-2-yl,furan-2-yl, 1,2,4-triazin-3-yl, 5,6-dimethyl-1,2,4-triazin-3-yl,5,6-diphenyl-1,2,4-triazin-3-yl, 1,2,4-oxadiazol-3-yl,4H-1,2,4-triazol-3-yl, 5-thioxo-4,5-dihydro-1H-1,2,4-triazol-3-yl,4,5-dihydro-1H-imidazol-2-yl, 4-phenylthiazol-2-yl, 1H-tetrazol-5-yl,1,4,5,6-tetrahydropyrimidin-2-yl, or9-oxo-9H-indeno[1,2-e][1,2,4]triazin-3-yl;

or a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

Preferably, the present invention provides a compound of formula (I), inwhich R₁ is hydrogen, R₂ is (5-7) membered heteroaryl; R₃ is (C₁-C₄)alkyl, with the proviso that R₂ is not 1,3-oxazol-2-yl, furan-2-yl,1,2,4-triazin-3-yl, 5,6-dimethyl-1,2,4-triazin-3-yl,5,6-diphenyl-1,2,4-triazin-3-yl, 1,2,4-oxadiazol-3-yl,4H-1,2,4-triazol-3-yl, 5-thioxo-4,5-dihydro-1H-1,2,4-triazol-3-yl,4,5-dihydro-1H-imidazol-2-yl, 4-phenylthiazol-2-yl, 1H-tetrazol-5-yl,1,4,5,6-tetrahydropyrimidin-2-yl, or9-oxo-9H-indeno[1,2-e][1,2,4]triazin-3-yl;

or a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

In another aspect, the present invention provides a compound of formula(I), wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ isalkynyl; R₃ is hydrogen; or a pharmaceutically acceptable salt thereof;or an optical isomer thereof; or a mixture of optical isomers.Preferably, R₁ and R₃ are hydrogen, R₂ is (C₂-C₄) alkynyl. Alsopreferably, R₁ and R₃ are hydrogen, R₂ is ethynyl.

In one embodiment, the present invention provides the compounds offormula (I) or a pharmaceutically acceptable salt thereof; or an opticalisomer thereof; or a mixture of optical isomers, represented by thefollowing structures:

or a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.

The present invention provides for compounds of formula (I),pharmaceutical compositions employing such compounds comprising apharmaceutically acceptable salts thereof, or a pharmaceuticallyacceptable carrier/excipient thereof, and for methods of using suchcompounds.

Any asymmetric carbon atom on the compounds of the present invention canbe present in the (R)-, (S)- or (R,S)-configuration, preferably in the(R)- or (S)-configuration.

Any resulting mixtures of isomers can be separated on the basis of thephysicochemical differences of the constituents, into the pure geometricor optical isomers, diastereomers, racemates, for example, bychromatography and/or fractional crystallization.

Any resulting racemates of final products or intermediates can beresolved into the optical antipodes by known methods, e.g., byseparation of the diastereomeric salts thereof, obtained with anoptically active acid or base, and liberating the optically activeacidic or basic compound. In particular, the hydroxamide or sulfonamidemoiety may thus be employed to resolve the compounds of the presentinvention into their optical antipodes, e.g., by fractionalcrystallization of a metal (e.g., Zn²⁺) complex formed with an opticallyactive co-ligand, e.g., L- or D-histidine. Racemic products can also beresolved by chiral chromatography, e.g., high pressure liquidchromatography (HPLC) using a chiral adsorbent.

It will be appreciated by those skilled in the art that compounds of theinvention having a chiral center may exist in and be isolated inoptically active and racemic forms. Some compounds may exhibitpolymorphism. It is to be understood that the present inventionencompasses any racemic, optically-active, polymorphic, orstereoisomeric form, or mixtures thereof, of a compound of theinvention, which possess the useful properties described herein, itbeing well known in the art how to prepare optically active forms (forexample, by resolution of the racemic form by recrystallizationtechniques, by synthesis from optically-active starting materials, bychiral synthesis, or by chromatographic separation using a chiralstationary phase.

The compounds of the present invention are useful in inhibitingtumor/cancer cell growth or cell proliferation of tumor/cancer cells,slowing down cell cycle progression in tumor/cancer cells. In addition,the compounds of the present invention are shown to induce apoptosis.Induction of apoptosis has been used as an important chemotherapyapproach in treating cancertumor. Accordingly, the compounds of thepresent invention have valuable pharmaceutical properties, they can beuseful as anti-proliferation and anti-tumor/anti-cancer agents.

Therefore, in one aspect, the compounds of the present invention can beused for inhibiting cell proliferation both in vitro and in vivo. In oneembodiment, the compounds of the present invention can be used toinhibit cell proliferation in a tumor/cancer cell by contacting thetumor/cancer cell with an effective amount of said compounds. In oneembodiment, the compounds of the present invention can be used to treatcellular proliferation diseases or conditions. Said diseases caninclude, but are not limited to, cancer, dysplasias, neoplasias, skin ormucosal warts, autoimmune diseases, fungal disorders, arthritis, graftrejection, inflammatory bowel disease, cellular proliferation inducedafter medical procedures, including, but not limited to, surgery,angioplasty, and the like.

In another aspect, the compounds of the present invention can be usedfor inhibiting tumor/cancer growth both in vitro and in vivo. In oneembodiment, the compounds can be used for inhibiting tumor/cancer cellgrowth by contacting the tumor/cancer cell with an effective amount ofsaid compounds. In one embodiment, the invention provides a method ofusing the compounds of the present invention for inhibiting tumor orcancer growth. Tumors or cancers that are treatable according to themethods include, for example, tumors or cancers located in the breast,lung, thyroid, lymph node, genitourinary system, kidney, ureter,bladder, ovary, testis, prostate, musculoskeletal system, bone, skeletalmuscle, bone marrow, gastrointestinal tract, stomach, esophagus, smallbowel, colon, rectum, pancreas, liver, smooth muscle, central orperipheral nervous system, brain, spinal cord, nerves, head, neck, ear,eye, nasopharynx, oropharynx, salivary gland, cardiovascular system,oral cavity, tongue, larynx, hypopharynx, soft tissues, skin, cervix,anus, retina, and/or heart of a mammal.

In one embodiment the invention provides a method of using the compoundsof the present invention to treat a neoplastic disease, or atumor/cancer. As used herein, the term “neoplastic disease” refers toany abnormal growth of cells or tissues being either benign(non-cancerous) or malignant (cancerous). Neoplastic diseases that aretreatable according to the methods of the invention include, forexample, neoplasms from acute myelogenous leukemia, chronic lymphocyticleukemia, chronic myelogenous leukemia, cutaneous T-cell lymphoma,hairy-cell leukemia and non-Hodgkin's lymphoma.

Additionally, the present invention provides:

-   -   a compound of the present invention for use as a medicament;    -   use of a compound of the present invention for the preparation        of a medicament for inhibiting cell proliferation in        tumor/cancer cells, or slowing down cell cycle progression in        tumor/cancer cells;    -   use of a compound of the present invention for the preparation        of a medicament for treating cellular proliferation diseases or        conditions;    -   use of a compound of the present invention for the preparation        of a medicament for inhibiting tumor/cancer growth both in vitro        and in vivo;    -   use of a compound of the present invention for the preparation        of a medicament for treating a neoplastic disease.    -   use of a compound of the present invention for the preparation        of a medicament for treating a tumor or cancer.

Processes for preparing compounds of formula I are provided as furtherembodiments of the invention and are illustrated by the followingprocedures in which the meanings of the generic radicals are as givenabove unless otherwise qualified.

A compound of formula I can be prepared as follows.

Chemistry

Suzuki-Miyaura cross-coupling reactions of 7-iodotubericidine 1 (Scheme1, Table 1) {for preparation, see Seela, F.; Ming, X. Tetrahedron 2007,63, 9850-9861} with corresponding aryl and hetaryl boronic acidsperformed under Shaughnessy aqueous conditions provided desired7-substituted-7-deazaadenosines 2a-n.

TABLE 1 Suzuki cross-coupling reactions Entry R₂ R₁B(OH)₂ Product(yield) 1

2a (54%) 2

2b (36%) 3

2c (48%) 4

2d (47%) 5

2e (18%) 6

2f (35%) 7

2g (32%) 8

2h (28%) 9

2i (56%) 10

2j (27%) 11

2k (77%) 12

2l (63%) 13

2m (9%) 14

2n (82%)

It should be noted that N-protecting group in both starting pyrrolylboronic acids were removed under the conditions of coupling (entry 11,12). Due to poor yield of Suzuki reaction (entry 13), 4-pyrazolylderivative 2m was alternatively prepared by Stille reaction of 1 with1-dimethylsulfamoyl-4-tributylstannylpyrazole {for preparation, see US20040157892 A1} and subsequent removal of dimethylsulfamoyl group underacidic conditions (1M aq HCl) in 68% overall yield aftercrystallization.

Ethynyl derivative 2o was prepared by Sonogashira reaction of 1 (Scheme2) with trimethylsilylacetylene and protodesilylation of TMS-ethynylderivative 3 under basic conditions. Triazolyl derivative 2p wasprepared by copper-mediated [3+2] cycloaddition of this ethynylderivative 2o with trimethylsilyl azide {Jin, T.; Kamijo, S.; Yamamoto,Y. Eur. J. Org. Chem. 2004, 3789-3791}.

Imidazolyl and thiazolyl derivatives 2q-s were prepared by Negishi orStille cross-coupling reactions of per-O-silylated 7-iodotubericidine 4(Scheme 3, Table 2) with corresponding protected organometallic reagentsand subsequent acidic deprotection.

Reactions

TABLE 2 Cross couplings and deprotection Product Entry R R₁—M (Yield) 1

2q (76%) 2

2r (77%) 3

2s (13%)

Analogous 2′-C-methyl-7-substituted-7-deazapurine ribosides wereprepared by aqueous Suzuki cross-coupling reactions of2′-C-methyl-7-iodo-7-deazaadenosine 6 (Scheme 4) with boronic acidsaffording products 7a-i.

The synthesis of required starting 2′-C-methyl riboside 6 started withacid promoted glycosylation of 6-chloro-7-iodo-7-deazapurine 8 (Scheme5) by per-O-benzoyl-2-C-methyl-β-D-ribofuranose 9 affording protected2′-C-methyl riboside 10 in 48% yield. Heating of compound 10 withammonia provided desired free nucleoside 6 in 69% yield.

For the synthesis of 7-substituted-7-deazaadenosine 5′-O-triphosphatesand 5′-O-monophosphates Suzuki reactions of 7-iodo-7-deazaadenosine5′-O-triphosphate 11 (Scheme 6, Table 3) and 5′-O-monophosphate 12 withboronic acids were carried out affording triphosphates 13a-f andmonophosphates 14a-f. Required starting iodo tri- and monophospahtes 11and 12 were prepared by convenient phosphorylation of 7-iodotubericidine1.

TABLE 3 Cross-coupling reactions of 11, 12 Product from 11 Product from12 Entry R₂ (yield) (yield) 1

13a (46%) 14a (94%) 2

13b (25%) 14b (47%) 3

13c (27%) 14c (34%) 4

13d (53%) 14d (51%) 5

13e (37%) 14e (45%) 6

13f (67%) 14f (47%)Salts and Hydrates

The compositions of this invention optionally comprise salts of thecompounds herein, especially pharmaceutically acceptable non-toxic saltscontaining, for example, Na⁺, Li⁺, K⁺, Ca⁺² and Mg⁺². Such salts mayinclude those derived by combination of appropriate cations such asalkali and alkaline earth metal ions or ammonium and quaternary aminoions with an acid anion moiety, typically a carboxylic acid. Monovalentsalts are preferred if a water soluble salt is desired. Some salts maybe useful as intermediates for purifying compounds of formula I or forpreparing other salts.

Metal salts typically are prepared by reacting the metal hydroxide witha compound of this invention. Examples of metal salts which are preparedin this way are salts containing Li⁺, Na⁺, and K⁺. A less soluble metalsalt can be precipitated from the solution of a more soluble salt byaddition of the suitable metal compound. In addition, salts may beformed from acid addition of certain organic and inorganic acids, e.g.,HCl, HBr, H₂SO₄, H₃PO₄ or organic sulfonic acids, to basic centers,typically amines, or to acidic groups. Finally, it is to be understoodthat the compositions herein comprise compounds of the invention intheir un-ionized, as well as zwitterionic form, and combinations withstoichiometric amounts of water as in hydrates.

Also included within the scope of this invention are the salts of theparental compounds with one or more amino acids. Any of the amino acidsdescribed above are suitable, especially the naturally-occurring aminoacids found as protein components, although the amino acid typically isone bearing a side chain with a basic or acidic group, e.g., lysine,arginine or glutamic acid, or a neutral group such as glycine, serine,threonine, alanine, isoleucine, or leucine.

Pharmaceutical Formulations

The compounds of this invention are formulated with conventionalcarriers and excipients, which will be selected in accord with ordinarypractice. Tablets will contain excipients, glidants, fillers, bindersand the like. Aqueous formulations are prepared in sterile form, andwhen intended for delivery by other than oral administration generallywill be isotonic. All formulations will optionally contain excipientssuch as those set forth in the Handbook of Pharmaceutical Excipients(1986). Excipients include ascorbic acid and other antioxidants,chelating agents such as EDTA, carbohydrates such as dextrin,hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid and thelike. The pH of the formulations ranges from about 3 to about 11, but isordinarily about 7 to 10.

While it is possible for the active ingredients to be administered aloneit may be preferable to present them as pharmaceutical formulations. Theformulations, both for veterinary and for human use, of the inventioncomprise at least one active ingredient, as above defined, together withone or more acceptable carriers therefore and optionally othertherapeutic ingredients. The carrier(s) must be “acceptable” in thesense of being compatible with the other ingredients of the formulationand physiologically innocuous to the recipient thereof.

The formulations include those suitable for the foregoing administrationroutes. The formulations may conveniently be presented in unit dosageform and may be prepared by any of the methods well known in the art ofpharmacy. Techniques and formulations generally are found in Remington'sPharmaceutical Sciences (Mack Publishing Co., Easton, Pa.). Such methodsinclude the step of bringing into association the active ingredient withthe carrier which constitutes one or more accessory ingredients. Ingeneral the formulations are prepared by uniformly and intimatelybringing into association the active ingredient with liquid carriers orfinely divided solid carriers or both, and then, if necessary, shapingthe product.

Formulations of the present invention suitable for oral administrationmay be presented as discrete units such as capsules, cachets or tabletseach containing a predetermined amount of the active ingredient; as apowder or granules; as a solution or a suspension in an aqueous ornon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion. The active ingredient may also beadministered as a bolus, electuary or paste.

A tablet is made by compression or molding, optionally with one or moreaccessory ingredients. Compressed tablets may be prepared by compressingin a suitable machine the active ingredient in a free-flowing form suchas a powder or granules, optionally mixed with a binder, lubricant,inert diluent, preservative, surface active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered active ingredient moistened with an inert liquid diluent. Thetablets may optionally be coated or scored and optionally are formulatedso as to provide slow or controlled release of the active ingredienttherefrom.

For administration to the eye or other external tissues e.g., mouth andskin, the formulations are preferably applied as a topical ointment orcream containing the active ingredient(s) in an amount of, for example,0.075 to 20% w/w (including active ingredient(s) in a range between 0.1%and 20% in increments of 0.1% w/w such as 0.6% w/w, 0.7% w/w, etc.),preferably 0.2 to 15% w/w and most preferably 0.5 to 10% w/w. Whenformulated in an ointment, the active ingredients may be employed witheither a paraffinic or a water-miscible ointment base. Alternatively,the active ingredients may be formulated in a cream with an oil-in-watercream base.

If desired, the aqueous phase of the cream base may include, forexample, at least 30% w/w of a polyhydric alcohol, i.e. an alcoholhaving two or more hydroxyl groups such as propylene glycol, butane1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol(including PEG 400) and mixtures thereof. The topical formulations maydesirably include a compound which enhances absorption or penetration ofthe active ingredient through the skin or other affected areas. Examplesof such dermal penetration enhancers include dimethyl sulphoxide andrelated analogs.

The oily phase of the emulsions of this invention may be constitutedfrom known ingredients in a known manner. While the phase may comprisemerely an emulsifier (otherwise known as an emulgent), it desirablycomprises a mixture of at least one emulsifier with a fat or an oil orwith both a fat and an oil. Preferably, a hydrophilic emulsifier isincluded together with a lipophilic emulsifier which acts as astabilizer. It is also preferred to include both an oil and a fat.Together, the emulsifier(s) with or without stabilizer(s) make up theso-called emulsifying wax, and the wax together with the oil and fatmake up the so-called emulsifying ointment base which forms the oilydispersed phase of the cream formulations.

Emulgents and emulsion stabilizers suitable for use in the formulationof the invention include Tween® 60, Span® 80, cetostearyl alcohol,benzyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodiumlauryl sulfate.

The choice of suitable oils or fats for the formulation is based onachieving the desired cosmetic properties. The cream should preferablybe a non-greasy, non-staining and washable product with suitableconsistency to avoid leakage from tubes or other containers. Straight orbranched chain, mono- or dibasic alkyl esters such as di-isoadipate,isocetyl stearate, propylene glycol diester of coconut fatty acids,isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate,2-ethylhexyl palmitate or a blend of branched chain esters known asCrodamol CAP may be used, the last three being preferred esters. Thesemay be used alone or in combination depending on the propertiesrequired. Alternatively, high melting point lipids such as white softparaffin and/or liquid paraffin or other mineral oils are used.

Pharmaceutical formulations according to the present invention compriseone or more compounds of the invention together with one or morepharmaceutically acceptable carriers or excipients and optionally othertherapeutic agents. Pharmaceutical formulations containing the activeingredient may be in any form suitable for the intended method ofadministration. When used for oral use for example, tablets, troches,lozenges, aqueous or oil suspensions, dispersible powders or granules,emulsions, hard or soft capsules, syrups or elixirs may be prepared.Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsincluding sweetening agents, flavoring agents, coloring agents andpreserving agents, in order to provide a palatable preparation. Tabletscontaining the active ingredient in admixture with non-toxicpharmaceutically acceptable excipient which are suitable for manufactureof tablets are acceptable. These excipients may be, for example, inertdiluents, such as calcium or sodium carbonate, lactose, lactosemonohydrate, croscarmellose sodium, povidone, calcium or sodiumphosphate; granulating and disintegrating agents, such as maize starch,or alginic acid; binding agents, such as cellulose, microcrystallinecellulose, starch, gelatin or acacia; and lubricating agents, such asmagnesium stearate, stearic acid or talc. Tablets may be uncoated or maybe coated by known techniques including microencapsulation to delaydisintegration and adsorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearatealone or with a wax may be employed.

Formulations for oral use may be also presented as hard gelatin capsuleswhere the active ingredient is mixed with an inert solid diluent, forexample calcium phosphate or kaolin, or as soft gelatin capsules whereinthe active ingredient is mixed with water or an oil medium, such aspeanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the invention contain the active materials inadmixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include a suspending agent, such as sodiumcarboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia,and dispersing or wetting agents such as a naturally occurringphosphatide (e.g., lecithin), a condensation product of an alkyleneoxide with a fatty acid (e.g., polyoxyethylene stearate), a condensationproduct of ethylene oxide with a long chain aliphatic alcohol (e.g.,heptadecaethyleneoxycetanol), a condensation product of ethylene oxidewith a partial ester derived from a fatty acid and a hexitol anhydride(e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension mayalso contain one or more preservatives such as ethyl or n-propylp-hydroxy-benzoate, one or more coloring agents, one or more flavoringagents and one or more sweetening agents, such as sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient ina vegetable oil, such as arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oral suspensionsmay contain a thickening agent, such as beeswax, hard paraffin or cetylalcohol. Sweetening agents, such as those set forth above, and flavoringagents may be added to provide a palatable oral preparation. Thesecompositions may be preserved by the addition of an antioxidant such asascorbic acid.

Dispersible powders and granules of the invention suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Suitable dispersing orwetting agents and suspending agents are exemplified by those disclosedabove. Additional excipients, for example sweetening, flavoring andcoloring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, a mineral oil, such as liquid paraffin, ora mixture of these. Suitable emulsifying agents includenaturally-occurring gums, such as gum acacia and gum tragacanth,naturally occurring phosphatides, such as soybean lecithin, esters orpartial esters derived from fatty acids and hexitol anhydrides, such assorbitan monooleate, and condensation products of these partial esterswith ethylene oxide, such as polyoxyethylene sorbitan monooleate. Theemulsion may also contain sweetening and flavoring agents. Syrups andelixirs may be formulated with sweetening agents, such as glycerol,sorbitol or sucrose. Such formulations may also contain a demulcent, apreservative, a flavoring or a coloring agent.

The pharmaceutical compositions of the invention may be in the form of asterile injectable preparation, such as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according tothe known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butane-diol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain approximately 1 to 1000 mg of active material compounded with anappropriate and convenient amount of carrier material which may varyfrom about 5 to about 95% of the total compositions (weight:weight). Thepharmaceutical composition can be prepared to provide easily measurableamounts for administration. For example, an aqueous solution intendedfor intravenous infusion may contain from about 3 to 500 μg of theactive ingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr can occur.

Formulations suitable for administration to the eye include eye dropswherein the active ingredient is dissolved or suspended in a suitablecarrier, especially an aqueous solvent for the active ingredient. Theactive ingredient is preferably present in such formulations in aconcentration of 0.5 to 20%, advantageously 0.5 to 10% particularlyabout 1.5% w/w.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the active ingredient in asuitable liquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for intrapulmonary or nasal administration have aparticle size for example in the range of 0.1 to 500 microns (includingparticle sizes in a range between 0.1 and 500 microns in incrementsmicrons such as 0.5, 1, 30 microns, 35 microns, etc.), which isadministered by rapid inhalation through the nasal passage or byinhalation through the mouth so as to reach the alveolar sacs. Suitableformulations include aqueous or oily solutions of the active ingredient.Formulations suitable for aerosol or dry powder administration may beprepared according to conventional methods and may be delivered withother therapeutic agents such as compounds heretofore used in thetreatment or prophylaxis of cancerous infections as described below.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents.

The formulations are presented in unit-dose or multi-dose containers,for example sealed ampoules and vials, and may be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example water for injection, immediatelyprior to use. Extemporaneous injection solutions and suspensions areprepared from sterile powders, granules and tablets of the kindpreviously described. Preferred unit dosage formulations are thosecontaining a daily dose or unit daily sub-dose, as herein above recited,or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the ingredients particularlymentioned above the formulations of this invention may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavoring agents.

The invention further provides veterinary compositions comprising atleast one active ingredient as above defined together with a veterinarycarrier therefore.

Veterinary carriers are materials useful for the purpose ofadministering the composition and may be solid, liquid or gaseousmaterials which are otherwise inert or acceptable in the veterinary artand are compatible with the active ingredient. These veterinarycompositions may be administered orally, parenterally or by any otherdesired route.

Compounds of the invention can also be formulated to provide controlledrelease of the active ingredient to allow less frequent dosing or toimprove the pharmacokinetic or toxicity profile of the activeingredient. Accordingly, the invention also provided compositionscomprising one or more compounds of the invention formulated forsustained or controlled release.

Effective dose of active ingredient depends at least on the nature ofthe condition being treated, toxicity, whether the compound is beingused prophylactically (lower doses) or against an active cancerousinfection, the method of delivery, and the pharmaceutical formulation,and will be determined by the clinician using conventional doseescalation studies. It can be expected to be from about 0.0001 to about100 mg/kg body weight per day. Typically, from about 0.01 to about 10mg/kg body weight per day. More typically, from about 0.01 to about 5mg/kg body weight per day. More typically, from about 0.05 to about 0.5mg/kg body weight per day. For example, the daily candidate dose for anadult human of approximately 70 kg body weight will range from 1 mg to1000 mg, preferably between 5 mg and 500 mg, and may take the form ofsingle or multiple doses.

Routes of Administration

One or more compounds of the invention (herein referred to as the activeingredients) are administered by any route appropriate to the conditionto be treated. Suitable routes include oral, rectal, nasal, topical(including buccal and sublingual), vaginal and parenteral (includingsubcutaneous, intramuscular, intravenous, intradermal, intrathecal andepidural), and the like. It will be appreciated that the preferred routemay vary with for example the condition of the recipient. An advantageof the compounds of this invention is that they are orally bioavailableand can be dosed orally.

Combination Therapy

Active ingredients of the invention are also used in combination withother active ingredients. Such combinations are selected based on thecondition to be treated, cross-reactivities of ingredients andpharmaco-properties of the combination. For example, when treatingcancer, the compositions of the invention can be combined with otherchemotherapeutic agents. The second chemotherapeutic agent can be anysuitable compound that has biological activity against one or more formsof cancer.

It is also possible to combine any compound of the invention with one ormore other active ingredients in a unitary dosage form for simultaneousor sequential administration to an cancer patient. The combinationtherapy may be administered as a simultaneous or sequential regimen.When administered sequentially, the combination may be administered intwo or more administrations. Second and third active ingredients in thecombination may have chemotherapeutic activity and include any of theadditional chemotherapeutic agents described herein. Exemplary activeingredients to be administered in combination with compounds of theinvention are described below.

Suitable additional chemotherapeutic agents include, e.g., antracyclines(e.g., doxorubicin, daunorubicin, epirubicin, idarubicin, andmitoxantrone); (b) other DNA intercalators (e.g., actinomycins C, D, B,etc.; podophyllotoxins, and epipodophyllatoxins (etoposide, teniposide,ctoposide)); (c) alkylating agents (e.g., mechlorethamine, melphalan,cyclophosphamide, chlorambucil, ifosfamide, carmustine, lomustine,busulfan, dacarbazine, cisplatin, carboplatin, oxaliplatin, iproplatin,and tetraplatin); (d) hormonal agents (e.g., antiestrogens estrogenantagonists (tamoxifen and other SERMs); LHRH agonists and antagonists(leuprolide acetate, goserelin, abarelix); aromatase inhibitors; andantiandrogens; (e) chemoprevention agents (e.g., NSAIDs andcis-retinoids); and (f) cell-cycle chemopreventative agents.

Alternatively, the additional chemotherapeutic agent can include, e.g.,antineoplasts. Representative antineoplasts include, e.g., adjuncts(e.g., levamisole, gallium nitrate, granisetron, sargramostimstrontium-89 chloride, filgrastim, pilocarpine, dexrazoxane, andondansetron); androgen inhibitors (e.g., flutamide and leuprolideacetate); antibiotic derivatives (e.g., doxorubicin, bleomycin sulfate,daunorubicin, dactinomycin, and idarubicin); antiestrogens (e.g.,tamoxifen citrate, analogs thereof, and nonsteroidal antiestrogens suchas toremifene, droloxifene and roloxifene); antimetabolites (e.g.,fludarabine phosphate, interferon alfa-2b recombinant, methotrexatesodium, plicamycin, mercaptopurine, and thioguanine); cytotoxic agents(e.g., doxorubicin, carmustine [BCNU], lomustine [CCNU], cytarabine USP,cyclophosphamide, estramucine phosphate sodium, altretamine,hydroxyurea, ifosfamide, procarbazine, mitomycin, busulfan,cyclophosphamide, mitoxantrone, carboplatin, cisplatin, interferonalfa-2a recombinant, paclitaxel, teniposide, and streptozocin); hormones(e.g., medroxyprogesterone acetate, estradiol, megestrol acetate,octreotide acetate, diethylstilbestrol diphosphate, testolactone, andgoserelin acetate); immunomodulators (e.g., aldesleukin); nitrogenmustard derivatives (e.g., melphalan, chlorambucil, mechlorethamine, andthiotepa) and steroids (betamethasone sodium phosphate and betamethasoneacetate).

Suitable additional chemotherapeutic agents include, e.g., alkylatingagents, antimitotic agents, plant alkaloids, biologicals, topoisomeraseI inhibitors, topoisomerase II inhibitors, and synthetics.

Representative alkylating agents include, e.g., asaley, AZQ, BCNU,busulfan, bisulphan, carboxyphthalatoplatinum, CBDCA, CCNU, CHIP,chlorambucil, chlorozotocin, cis-platinum, clomesone,cyanomorpholinodoxorubicin, cyclodisone, cyclophosphamide,dianhydrogalactitol, fluorodopan, hepsulfam, hycanthone, iphosphamide,melphalan, methyl CCNU, mitomycin C, mitozolamide, nitrogen mustard,PCNU, piperazine, piperazinedione, pipobroman, porfiromycin,spirohydantoin mustard, streptozotocin, teroxirone, tetraplatin,thiotepa, triethylenemelamine, uracil nitrogen mustard, and Yoshi-864.

Representative antimitotic agents include, e.g., allocolchicine,Halichondrin B, colchicine, colchicine derivatives, dolastatin 10,maytansine, rhizoxin, paclitaxel derivatives, paclitaxel,thiocolchicine, trityl cysteine, vinblastine sulfate, and vincristinesulfate.

Representative plant alkaloids include, e.g., actinomycin D, bleomycin,L-asparaginase, idarubicin, vinblastine sulfate, vincristine sulfate,mitramycin, mitomycin, daunorubicin, VP-16-213, VM-26, navelbine andtaxotere.

Representative biologicals include, e.g., alpha interferon, BCG, G-CSF,GM-CSF, and interleukin-2.

Representative topoisomerase I inhibitors include, e.g., camptothecin,camptothecin derivatives, and morpholinodoxorubicin.

Representative topoisomerase II inhibitors include, e.g., mitoxantron,amonafide, m-AMSA, anthrapyrazole derivatives, pyrazoloacridine,bisantrene HCL, daunorubicin, deoxydoxorubicin, menogaril, N, N-dibenzyldaunomycin, oxanthrazole, rubidazone, VM-26 and VP-16.

Representative synthetics include, e.g., hydroxyurea, procarbazine,o,p′-DDD, dacarbazine, CCNU, BCNU, cis-diamminedichloroplatimun,mitoxantrone, CBDCA, levamisole, hexamethylmelamine, all-trans retinoicacid, gliadel and porfimer sodium.

Alternatively, the additional chemotherapeutic agent can includetubulin-binding drugs and drugs that affect tubulin dynamics andfunction. This includes a variety of drugs that are chemically unrelatedto vinca alkaloids and taxanes (e.g. CP-248 [a derivative of exisulind]and ILX-651). These drugs have distinctive effects on cells at G2M-phaseand may have functionally independent effects on cells in G1 and or Sphase.

Alternatively, the additional chemotherapeutic agent can includeselective apoptotic anti-cancer drugs (SAANDs), which include sulindac,aptosyn, CP-461, CP-248 and related sulindac derived compounds thatinhibit one or more of the following isozymes of cyclic GMPphosphodiesterase (cGMP PDE): 1, 2, 5.

Alternatively, the additional chemotherapeutic agent can include drugsthat inhibit proteosomes (bortezomib or Velcade). Proteosomes degrademany ubiquitinated proteins that have been marked for activedestruction. Ubiquitinated proteins include many critical cell cycleregulatory molecules and molecules that regulate apoptosis at specificstages of the cell cycle. While proteosomes may degrade proteinsthroughout the cell cycle, the proteins that are degraded by proteosomesinclude some of the most critical cell cycle regulatory proteins. Theso-called “cell cycle active rationale” may be applied to the treatmentof diseases in various categories, including cancer,inflammatory/autoimmune diseases, and neurological diseases that involvedisorderly cell cycle and/or apoptosis.

Alternatively, the additional chemotherapeutic agent can include drugsthat inhibit heat shock protein 90 (HSP90), a ‘chaperonin’ thatparticipates in the degradation of ‘client’ proteins in the ubiquitinmediated proteosome pathway. Several drugs seem to exert theirantitumour effect by inhibiting the intrinsic ATPase activity of HSP90,resulting in degradation of HSP90 “client proteins” via the ubiquitinproteosome pathway. Examples include: geldanamycin, 17-allylaminogeldanamycin, 17-demethoxygeldanamycin and radicicol.

Suitable cell-cycle dependent biological agents or schedule-dependentbiological agents include drugs, proteins or other molecules that block,impede, or otherwise interfere with, cell cycle progression at theG1-phase, G1/S interface, S-phase, G2M interface, or M-phase of the cellcycle. These drugs are cell cycle-dependent or schedule-dependent.

Specifically, suitable cell-cycle dependent biological agents orschedule-dependent biological agents include:

(1) Analogues of uridine nucleosides, analogues of thymidinenucleosides, and analogues of uridine and thymidine nucleosides. Thesecompounds act at the S-phase in tumor cells, and possibly neovascularendothelial cells. These compounds include, e.g., 5-fluorodeoxyuridine(floxuridine, FUDR); 5-fluorouracil (5-FU); prodrugs of 5-FU (e.g.capecitabine, 5′-deoxy-5-fluorouridine, ftorafur, flucytosine);bromodeoxyuridine; and iododexoyuridine.

(2) Modulators of fluoropyrimidines. These compounds act at the S-phasein tumor cells, and possibly neovascular endothelial cells. Thesecompounds include, e.g., leurovorin, methotrexate and other folates;levamisole; acivicin; phosphonacetyl-L-aspartic acid (PALA); brequinar;5-ethynyluracil; and uracil.

(3) Cytidine analogues and cytidine nucleoside analogues. Thesecompounds act at the S-phase in tumor cells, and possibly neovascularendothelial cells. These compounds include, e.g., cytarabine (Ara-C,cytosine arabinoside); gemcitabine (2′,2′-difluorodeoxycytidine); and5-azacytidine.

(4) Purine analogues and purine nucleoside analogues. These compoundsact at the S-phase in tumor cells, and possibly neovascular endothelialcells. These compounds include, e.g., 6-thioguanine; 6-mercaptopurine;azathioprine; adenosine arabinoside (Ara-A);2′,2′-difluorodeoxyguanosine; deoxycoformycin (pentostatin); cladribine(2-chlorodeoxyadenosine); and inhibitors of adenosine deaminase.

(5) Antifolates. These compounds act at the S-phase in tumor cells, andpossibly neovascular endothelial cells. These compounds include, e.g.,methotrexate; aminopterin; trimetrexate; edatrexate;N10-propargyl-5,8-dideazafolic acid (CB3717); ZD1694,5,8-dideazaisofolic acid (IAHQ); 5,10-dideazatetrahydrofolic acid(DDATHF); 5-deazafolic acid (efficient substrate for FPGS); PT523 (Nalpha-(4-amino-4-deoxypteroyl)-N delta-hemiphthaloyl-L-ornithine);10-ethyl-10-deazaaminopterin (DDATHF, lomatrexol); piritrexim; 10-EDAM;ZD1694; GW1843; PDX (10-propargyl-10-deazaaminopterin); multi-targetedfolate (i.e. LY231514, permetrexed); any folate-based inhibitor ofthymidylate synthase (TS); any folate-based inhibitor of dihydrofolatereductase (DHFR); any folate-based inhibitor of glycinamideribonucleotide transformylase (GARTF); any inhibitor offolylpolyglutamate synthetase (FPGS); and any folate-based inhibitor ofGAR formyl transferase (AICAR transformylase).

(6) Other antimetabolites. These compounds act at the S-phase in tumorcells, and possibly neovascular endothelial cells. These compoundsinclude, e.g., hydroxyurea and polyamines.

(7) S-phase specific radiotoxins (deoxythymidine analogues). Thesecompounds act at the S-phase in all cells undergoing DNA synthesis. Thecompounds are incorporated into chromosomal DNA during S-phase. Thesecompounds include, e.g., [125I]-iododeoxyuridine;[123I]-iododeoxyuridine; [124I]-iododeoxyuridine;[80mBr]-iododeoxyuridine; [131I]-iododeoxyuridine; and[211At]-astatine-deoxyuridine.

(8) Inhibitors of enzymes involved in deoxynucleosidedeoxynucleotidemetabolism. These compounds act at the S-phase in tumor cells, andpossibly neovascular endothelial cells. These compounds include, e.g.,inhibitors of thymidylate synthase (TS); inhibitors of dihydrofolatereductase (DHFR); inhibitors of glycinamide ribonucleotidetransformylase (GARTF); inhibitors of folylpolyglutamate synthetase(FPGS); inhibitors of GAR formyl transferase (AICAR transformylase);inhibitors of DNA polymerases (DNA Pol; e.g. aphidocolin); inhibitors ofribonucleotide reductase (RNR); inhibitors of thymidine kinase (TK); andinhibitors of topoisomerase I enzymes (e.g. camptothecins, irinotecan[CPT-11, camptosar], topotecan, NX-211 [lurtotecan], rubitecan, etc.).

(9) DNA chain-terminating nucleoside analogues. These compounds actspecifically on S-phase cells and are incorporated into chromosomal DNAduring S-phase; terminate growing DNA strand. These compounds include,e.g., acyclovir; abacavir; valacyclovir; zidovudine (AZT); didanosine(ddI, dideoxycytidine); zalcitabine (ddC); stavudine (D4T); lamivudine(3TC); Any 2′ 3′-dideoxy nucleoside analogue; and any 2′ 3′-dideoxynucleoside analogue that terminates DNA synthesis. These compoundsinclude, e.g., inhibitors of growth factor receptor tyrosine kinasesthat regulate progression through the G1-phase, G1/S interface, orS-phase of the cell cycle (e.g. EGF receptors, HER-2 neu/c-erbB2receptor, PDGF receptors, etc; [e.g. trastusumab, iressa, erbitux,tarceva]); inhibitors of non-receptor tyrosine kinases (e.g. c-srcfamily of tyrosine kinases; [e.g. Gleevec]); inhibitors ofserine-threonine kinases that regulate progression through the G1-phase,G1/S interface or S-phase of the cell cycle (e.g. G1 cyclin-dependentkinases, G1/S cyclin-dependent kinases, and S cyclin-dependent kinases[e.g. CDK2, CDK4, CDK5, CDK6]; mitogen-activated kinases; MAP kinasesignaling pathway); inhibitors of G1-phase, G1/S interface or S-phasecyclins [e.g. cyclins D1, D2, D3, E, and A]); inhibitors of G-proteinsand cGMP phosphodiesterases that positively regulate cell cycleprogression at the G1-phase, G1/S interface or S-phase of the cellcycle; drugs that inhibit the induction of immediate early responsetranscription factors (e.g. N-terminal c-jun kinase, c-myc); and drugsthat inhibit proteosomes that degrade ‘negative’ cell cycle regulatorymolecules (e.g. p53, p27Kip1; [e.g. bortezomib]).

(10) Cytokines, growth factors, anti-angiogenic factors and otherproteins that inhibit cell cycle progression at the G1-phase or G1/Sinterface of the cell cycle. These compounds act at G1, G1/S or S-phaseof the cell cycle in tumor cells, and in some cases, neovascularendothelial cells. These compounds include, e.g., interferons;interleukins; somatostatin and somatostatin analogues (octreotide,sandostatin LAR); and many anti-angiogenic factors inhibit cellproliferation of endothelial cells at the G1 or G1/S phases of the cellcycle.

(11) Drugs and compounds that inhibit cell cycle progression at the G2Minterface, or M-phase of the cell cycle. These compounds act at G2Minterface or M-phase of the cell cycle in tumor cells, and in somecases, neovascular endothelial cells. These compounds include, e.g., (a)microtubule-targeting drugs taxanes (e.g., taxol, taxotere, epothilones,and other taxanes and derivatives); (b) microtubule-targeting drugsvinca alkaloids (e.g., vinblastine, vincristine, vindesine; vinflunine,vinorelbine, vinzolidine, nocadazole, and colchicines); (c)microtubule-targeting drugs others (e.g., estramustine, CP-248 andCP-461); (d) inhibitors of serine-threonine kinases that regulateprogression through the G2M interface or M-phase of the cell cycle(e.g., inhibitors of G2M cyclin-dependent kinases (e.g. CDC2);inhibitors of M-phase cyclins (e.g. cyclin B) and any drug that blocks,impedes, or otherwise interferes with, cell cycle progression at the G2Minterface, or M-phase of the cell cycle).

(12) Radiopharmaceuticals useful in radiation therapy and/or diagnosis.A suitable class of radioisotopes decays by a nuclear disintegrationprocess known as the “Auger Process” or “Auger Cascade”. Auger emittingisotopes generate short acting electrons that efficiently cleave duplexDNA. Suitable Auger-emitting radionuclides include, e.g., 125-Iodine,123-Iodine and 80m-Bromine. Suitable corresponding halogenatedpryimidine and purine nucleosides include, e.g.,5-125Iodo-2′-deoxyuridine, 5-123Iodo-2′-deoxyuridine,5-80mBromo-2′-deoxyuridine and 8-80mBromo-2′-guanidine.

Growth Factors

Many growth factors and cytokines have the capacity to stimulatemalignant cells to traverse specific points in the cell cycle. Forexample, G-CSF or GM-CSF can stimulate leukemic blasts in acute myeloidleukemia to traverse the G1/S interface. This increases the cells'susceptibility to cell-cycle specific drugs, such as cytarabine. Similarstrategies have been tested using EGF and cytotoxic drugs for solidtumors. In order to respond the growth factor, cells must be at aspecific stage of the cell cycle, e.g., at the G1/S interface. Thecontinuous presence of a growth factor could be beneficial, because atany given time, only a subset of the blasts are at G1S. Thus, the growthfactors act in a cell cycle specific fashion. Similar logic can beapplied to the use of hematopoietic growth factors used to treatneutropenia, anemia and thrombocytopenia.

As such, peptide protein growth factors can be employed in the presentinvention to promote survival of normal non-malignant cell lineages. Onebenefit in using such substances is the ability to protect proliferatingcells in bone marrow, skin, oral and gastrointestinal mucosa, and hairfollicles.

Examples of substances within this category include, e.g., hematopoieticgrowth factors: G-CSF, GM-CSF, erythropoietin, thrombopoietin andbiologically active derivatives of these peptides; keratinocyte growthfactor (KGF) for mucositis; B-lymphocyte stimulating pepdie (BLys);platelet derived growth factor (PDGF), epithelial growth factor (EGF),TGF-alpha and related growth factors; interleukins (e.g. IL-2, IL-6);other cytokines, growth factors and peptides that stimulateproliferation of non-malignant cells that need to be protected.

Therapeutic Growth Factors/Cytokines

Some therapeutic growth factors/cytokines can inhibit cell proliferationof cancer cells and/or neovascular cells at specific stages of the cellcycle. For example, interferons, somatostatin, octreotide and analoguesthereof, thrombospondin and troponin-I inhibit neovascular endothelialcell proliferation by reducing the rate at which the cells enterS-phase. As such, any one or more of these substances can be employed inthe present invention.

The combination therapy may provide “synergy” and “synergistic effect”,i.e. the effect achieved when the active ingredients used together isgreater than the sum of the effects that results from using thecompounds separately. A synergistic effect may be attained when theactive ingredients are: (1) co-formulated and administered or deliveredsimultaneously in a combined formulation; (2) delivered by alternationor in parallel as separate formulations; or (3) by some other regimen.When delivered in alternation therapy, a synergistic effect may beattained when the compounds are administered or delivered sequentially,e.g., in separate tablets, pills or capsules, or by different injectionsin separate syringes. In general, during alternation therapy, aneffective dosage of each active ingredient is administered sequentially,i.e. serially, whereas in combination therapy, effective dosages of twoor more active ingredients are administered together.

Metabolites of the Compounds of the Invention

Also falling within the scope of this invention are the in vivometabolic products of the compounds described herein. Such products mayresult for example from the oxidation, reduction, hydrolysis, amidation,esterification, phosphorylation and the like of the administeredcompound, primarily due to enzymatic processes. Accordingly, theinvention includes compounds produced by a process comprising contactinga compound of this invention with a mammal for a period of timesufficient to yield a metabolic product thereof. Such products typicallyare identified by preparing a radiolabelled (e.g., C¹⁴ or H³) compoundof the invention, administering it to cells culturing in vitro orparenterally in a detectable dose (e.g., greater than about 0.5 mg/kg)to an animal such as rat, mouse, guinea pig, monkey, or to man, allowingsufficient time for metabolism to occur (typically about 30 seconds to30 hours) and isolating its conversion products from the urine, blood orother biological samples. These products are easily isolated since theyare labeled (others are isolated by the use of antibodies capable ofbinding epitopes surviving in the metabolite). The metabolite structuresare determined in conventional fashion, e.g., by MS or NMR analysis. Ingeneral, analysis of metabolites is done in the same way as conventionaldrug metabolism studies well-known to those skilled in the art. Theconversion products, so long as they are not otherwise found in vivo,are useful in diagnostic assays for therapeutic dosing of the compoundsof the invention even if they possess no anti-cancer activity of theirown.

Recipes and methods for determining stability of compounds in surrogategastrointestinal secretions are known. Compounds are defined herein asstable in the gastrointestinal tract where less than about 50 molepercent of the protected groups are deprotected in surrogate intestinalor gastric juice upon incubation for 1 hour at 37° C. Simply because thecompounds are stable to the gastrointestinal tract does not mean thatthey cannot be hydrolyzed in vivo.

In one embodiment of the invention, the compound is in an isolated andpurified form. Generally, the term “isolated and purified” means thatthe compound is substantially free from biological materials (e.g.blood, tissue, cells, etc.). In one specific embodiment of theinvention, the term means that the compound or conjugate of theinvention is at least about 50 wt. % free from biological materials; inanother specific embodiment, the term means that the compound orconjugate of the invention is at least about 75 wt. % free frombiological materials; in another specific embodiment, the term meansthat the compound or conjugate of the invention is at least about 90 wt.% free from biological materials; in another specific embodiment, theterm means that the compound or conjugate of the invention is at leastabout 98 wt. % free from biological materials; and in anotherembodiment, the term means that the compound or conjugate of theinvention is at least about 99 wt. % free from biological materials. Inanother specific embodiment, the invention provides a compound orconjugate of the invention that has been synthetically prepared (e.g.,ex vivo).

The anti-cancer activity of a compound may be determined usingpharmacological models which are well known to the art, for exampleusing Test A or B described below.

Test A: Cytostatic Cell Culture Assay (CC₅₀) for Solid Tumor Cell Lines

This assay is based on quantification of cell mass determination by acolorimetric detection of the cell associated proteins. The assay relieson the ability of sulforhodamine B (SRB) to bind to protein componentsof cells that have been fixed to tissue-culture plates bytrichloroacetic acid (TCA). SRB is a bright-pink aminoxanthene dye withtwo sulfonic groups that bind to basic amino-acid residues under mildacidic conditions, and dissociate under basic conditions. As the bindingof SRB is stochiometric, the amount of dye extracted from stained cellsis directly proportional to the cell mass.

Cell lines: All cell lines are obtained from ATCC (Manassas, Va.).Cultivation media containing Glutamax, and trypsin are purchased fromInvitrogen (Carlsbad, Calif.). Doxorubicin, Tubercidin, Clofarabine, TCAand SRB are from Sigma-Aldrich (St. Louis, Mo.). Gemcitabine is obtainedfrom Moravek Biochemicals (Brea, Calif.)

Assay Protocol:

-   1. Maintain cell lines in the media listed in Table 1. Trypsinize    the sub-confluent cells, count them, and adjust the cell    concentrations according to the cell counts listed in Table 1.-   2. Distribute the cells into the 96-well plates in 150 μL of media.    Incubate the plates overnight in humidified CO₂ incubator at 37° C.-   3. Fix one plate of each cell line with TCA. Discard the cultivation    media from the plates by flicking them gently and add 100 μL cold    10% (vol/vol) TCA to each well. Incubate the plates at 4 degree    refrigerator for 1 hour. Discard TCA from the plates by flicking    them gently. Rinse plates four times in a washing basin containing    tap water. Store the plates at room temperature. These plates    represent cell counts on day zero.-   4. Prepare a set of medium solutions containing various    concentrations of tested compounds by making 5-fold serial dilutions    in 96-well plate. Add 50 μL of the diluted compounds per well.    Include controls with untreated cells and cells treated with    doxorubicin, tubercidin, clofarabine and/or gemcitabine.-   5. Incubate the plates for 5 days at 37° C.-   6. Fix the plates with TCA. Discard the cultivation media from the    plates by flicking them gently and add 100 μL cold 10% (vol/vol) TCA    to each well. Incubate the plates at 4 degree refrigerator for 1    hour. Discard TCA from the plates by flicking them gently. Rinse    plates four times in a washing basin containing tap water.-   7. Remove excess water by tapping the plates face down, gently on a    paper towel. Allow the plates to air-dry at room temperature.-   8. Add 100 μL of 0.057% SRB solution in 1% (vol/vol) acetic acid to    each well of the plates fixed with TCA on day zero and five. Leave    at room temperature for 30 minutes.-   9. Flick the plates gently to discard SRB. Rinse the plates four    times with 1% (vol/vol) Acetic Acid.-   10. Store the plates at 37° C. incubator to facilitate faster    drying.-   11. Once the plates are completely dry, add 200 μL of 10 mM Tris    base solution (pH 10.5) to each well. Leave at room temperature for    30 minutes for SRB to solubilize.-   12. Measure the OD at 500 nm in a microplate reader.-   13. Calculate the percentage of cell-growth inhibition using the    next formula:    % of control cell growth=100×(OD _(sample)−mean OD _(day0))/(OD    _(neg control)−mean OD _(day0)).    For CC₅₀ determination, plot a dose-response curves between the    compound concentration and percent of growth inhibition. CC₅₀ values    can be derived by fitting dose-response curves using sigmoidal dose    response equation.

TABLE 4 Cultivation conditions for solid tumor cell lines SeedingDissociation CELL LINE Medium Density Agent HCT 116-Colon RPMI, 10% FBS, 800 cells/well Trypsin 1X Pen/Strep HCT 15-Colon RPMI, 10% FBS, 1600cells/well Trypsin 1X Pen/Strep BT549 RPMI, 10% FBS, 4000 cells/wellTryple Express 1X Pen/Strep (Invitrogen) HS 578-Breast RPMI, 10% FBS,4000 cells/well Tryple Express 1X Pen/Strep (Invitrogen) PC3-ProstateF12K, 10% FBS, 2500 cells/well Trypsin 1X Pen/Strep DU145-Prostate MEM,10% FBS,  800 cells/well Trypsin 1X Pen/Strep H23-Lung RPMI, 10% FBS,6000 cells/well Trypsin 1X Pen/Strep A549-Lung RPMI, 10% FBS, 1500cells/well Trypsin 1X Pen/StrepTest B: Cytostatic Cell Culture Assay (CC₅₀) for Lymphoid Tumor CellLines

This test is typically performed with cell lines that are derived fromhematological tumors and grow in suspension. An example of such cellline is human MT-4 T-lymphoid cell line used for the determination ofcytostatic activity of tested compounds. MT-4 cells were obtained fromthe NIH AIDS Research and Reference Reagents Program and were maintainedin RPMI-1640 medium supplemented with 10% FBS and antibiotics. Cellswere passaged in suspension twice a week and maintained at densitiesbelow 500,000 cells/mL For CC₅₀ determination, cells were seeded into384-well plates at 2,000 cells/well in 20 μL of culture medium.Compounds were serially diluted in culture medium and added intriplicate to a final assay volume of 40 μL/well. Plates were incubatedfor 5 days with tested compounds. At the end of incubation, cellviability was determined by addition of 40 μL of CellTiter Glo reagentfollowed by a luminescence read-out.

CC₅₀ values were determined as a concentration of each tested compoundresulting in 50% reduction in cell viability signal. Data analysis andCC₅₀ value calculations were done using GraphPad Prism software(GraphPad Software, San Diego, Calif.) by applying nonlinear regression.

Representative compounds of the invention typically have activityagainst one or more of the above cell lines with a CC₅₀ of less thanabout 20 μM. Some representative compounds of the invention haveactivity against one or more of the above cell lines with a CC₅₀ of lessthan about 1 μM. Still other representative compounds of the inventionhave activity against one or more of the above cell lines with a CC₅₀ ofless than about 0.1 μM.

Data for representative compounds of the invention from Tests A and Bare shown in the following Table 5.

TABLE 5 CC₅₀ (μM) against human tumor cell lines Lung Prostate ColonBreast T-lymphoid Geometric Compound A549 NCI H23 Du145 PC3 HCT116 HCT15HS578 BT549 MT-4 mean all 4-Amino-5-(1H-imidazol- 0.006 0.001 0.0020.001 0.049 0.0038 2-yl)-7-(β-D- ribofuranosyl)- 7H-pyrrolo[2,3-d]pyrimidine (2s) 4-Amino-7-(β-D- 0.005 0.042 0.093 0.015 0.042 0.0256ribofuranosyl)-5- (thiazol-2-yl)-7H- pyrrolo[2,3- d]pyrimidine (2q)4-Amino-5-(1H-imidazol- 0.002 0.002 0.003 0.002 0.028 0.00384-yl)-7-(β-D- ribofuranosyl)- 7H-pyrrolo[2,3- d]pyrimidine (2r)4-Amino-5-(1H-pyrazol- 0.035 0.015 0.018 0.008 0.018 0.01713-yl)-7-beta-D- ribofuranosyl)- 7H-pyrrolo[2,3- d]pyrimidine (2n)4-Amino-7-(β-D- 0.004 0.005 0.004 0.003 0.018 0.0052ribofuranosyl)-5-(1H- 1,2,3-triazol-4-yl)- 7H-pyrrolo[2,3- d]pyrimidine(2p) 4-Amino-5-ethynyl-7- 0.001 0.011 0.002 0.001 0.001 0.0019(β-D-ribofuranosyl)- 7H-pyrrolo[2,3- d]pyrimidine (2o)4-Amino-5-(1H-pyrrol- 0.1604 0.333 0.0878 0.0652 0.1129 0.1292 0.12832-yl)-7-(β-D- ribofuranosyl)- 7H-pyrrolo[2,3- d]pyrimidine (2k)4-Amino-5-(1H-pyrrol- 0.0053 0.033 0.0290 0.0059 0.0044 0.0080 0.0240.0114 3-yl)-7-(β-D- ribofuranosyl)- 7H-pyrrolo[2,3- d]pyrimidine (2l)4-Amino-5-(benzofuran- 7.02 5.40 6.29 3.74 6.73 >10 4.8 5.53852-yl)-7-(beta-D- ribofuranosyl)- 7H-pyrrolo[2,3- d]pyrimidine (2j)4-Amino-5-(1H-pyrazol- 0.192 0.144 0.014 0.970 0.012 0.050 0.110 0.08194-yl)-7-(beta-D- ribofuranosyl)- 7H-pyrrolo[2,3-d] pyrimidine (2m)4-Amino-7-(beta-D- 0.016 0.073 0.097 1.323 0.007 0.050 0.028 0.1240.0607 ribofuranosyl)- 5-(thiophen-3-yl)- 7H-pyrrolo[2,3- d]pyrimidine(2i) 4-Amino-5-(furan- 0.034 0.198 0.330 0.418 0.015 0.021 0.028 0.1700.0781 3-yl)-7-(beta-D- ribofuranosyl)- 7H-pyrrolo[2,3- d]pyrimidine(2h) 4-Amino-5-(naphtalen- >10 >10 >10 >10 >10 1-yl)-7-(beta-D-ribofuranosyl)- 7H-pyrrolo[2,3- d]pyrimidine (2d) 4-Amino-5-(naphtalen-5.35 1.05 5.67 2.16 2.8806 2-yl)-7-(beta-D- ribofuranosyl)-7H-pyrrolo[2,3- d]pyrimidine (2e) 4-Amino-5-[4-(methyl- 0.39 1.87 1.291.97 4.4 1.5235 thio)phenyl]-7-(beta- D-ribofuranosyl)- 7H-pyrrolo[2,3-d]pyrimidine (2c) 4-Amino-5-(4-methoxy- 0.701 0.856 0.152 1.106 0.6330.544 0.811 4.3 0.7683 phenyl)-7-(beta-D- ribofuranosyl)-7H-pyrrolo[2,3- d]pyrimidine (2b) 4-Amino-7-(beta-D- 0.0070 0.0037 0.1331.62 0.0026 0.011 0.012 0.035 0.0226 ribofuranosyl)- 5-(thiophen-2-yl)-7H-pyrrolo[2,3- d]pyrimidine (2g) 4-Amino-5-(furan-2- 0.035 0.294 0.0190.004 0.048 0.003 0.017 0.086 0.0256 yl)-7-(beta-D- ribofuranosyl)-7H-pyrrolo[2,3- d]pyrimidine (2f) Doxorubicine 0.016 0.005 0.021 0.0110.006 0.010 0.0101 Tubercidin 0.001 0.011 0.018 0.048 0.001 0.011 0.0980.021 0.0103 Clofarabine 0.086 0.040 0.125 0.063 0.106 0.180 1.241 0.0510.1158 Gemcitabine 0.007 0.002 0.003 0.006 0.002 0.003 0.001 0.001 0.0020.0024

Representative compounds of the invention are also found to inhibitadenosine kinase from Mycobacterium. Accordingly, in one embodiment, theinvention also provides a method for inhibiting an adenosine kinase(e.g. an adenosine kinase from Mycobacterium) comprising contacting theadenosine kinase with a compound of formula I or a pharmaceuticallyacceptable salt thereof.

In another embodiment, the invention also provides a method for treatinga disease associated with adenosine kinase activity in an animalcomprising administering to an animal (e.g. a mammal such as a human) inneed of such therapy, an effective adenosine kinase inhibiting amount ofa compound of formula I or a pharmaceutically acceptable salt thereof.Diseases associated with adenosine kinase activity may includeinflammation, sepsis, arthritis, rheumatoid arthritis, osteoarthritis,autoimmune diseases, burns, adult respiratory distress syndrome,inflammatory bowel syndrome, necrotizing enterocolitis, chronicobstructive pulmonary disease, psoriasis, conjunctivitis, iridocyclitis,ischemia, reperfusion injury, peripheral vascular disease, pancreatitis,atherosclerosis, meningitis, vasculitis, dermatitis, myositis, renalinflammation, sepsis, septicemia (e.g. endotoxemia), and septic shock(e.g. endotixic shock).

In another embodiment, the invention also provides a method for treatingtuberculosis in an animal (e.g. a mammal such as a human) comprisingadministering a compound of formula I or a pharmaceutically acceptablesalt thereof to the animal.

In another embodiment, the invention also provides the use of a compoundof formula I or a pharmaceutically acceptable salt thereof to prepare amedicament for inhibiting an adenosine kinase in an animal (e.g. amammal such as a human).

In another embodiment, the invention also provides the use of a compoundof formula I or a pharmaceutically acceptable salt thereof to prepare amedicament for treating a disease associated with adenosine kinaseactivity in an animal (e.g. a mammal such as a human).

In another embodiment, the invention also provides the use of a compoundof formula I or a pharmaceutically acceptable salt thereof to prepare amedicament for treating tuberculosis in an animal (e.g. a mammal such asa human).

Abbreviations

-   AcOEt ethylacetate-   Boc tert-butoxycarbonyl-   bd broad doublet-   bs broad singlet-   Bu butyl-   Bz benzoyl-   calcd calculated-   cat. catalyst-   d doublet-   dd doublet of doublets-   ddd doublet of doublet of doublets-   DMF dimethylformamide-   DMSO dimethylsulfoxide-   dt doublet of triplets-   Et ethyl-   EDTA ethylenediaminetetraacetic acid-   FAB fast atom bombardment-   gem geminal-   HPFC High performance flash chromatography-   HR high resolution-   i ipso-   iPr isopropyl-   IR infrared spectroscopy-   m multiplet-   m meta-   Me methyl-   MeCN acetonitrile-   MeOH methanol-   MeONa sodium methoxide-   MS mass spectrometry-   v wave number-   naphth naphthalenyl-   NMR nuclear magnetic resonance-   o ortho-   p para-   Ph phenyl-   PPh₃ triphenylphosphine-   Py pyridyl-   pyrr pyrrolyl-   q quartet-   rel. relative-   RT room temperature-   s singlet-   sat. saturated-   sol. solution-   t triplet-   TBS tert-butyldimethylsilyl-   td triplet of doublets-   THF tetrahydrofuran-   TFA trifluoroacetic acid-   TPPTS sodium triphenylphosphine trisulfonate-   Tr trityl, triphenylmethyl-   vic vicinal

The invention will now be illustrated by the following non-limitingExamples.

EXAMPLES

General Methods. Melting points were determined on a Kofler block.Optical rotations were measured at 25° C., [α]_(D) ²⁰ values are givenin 10⁻¹ deg cm² g⁻¹. NMR spectra were measured at 400 MHz for ¹H and100.6 MHz for ¹³C nuclei, at 500 MHz for ¹H and 125.8 MHz for ¹³C, or at600 MHz for ¹H and 151 MHz for ¹³C in CDCl₃ (TMS was used as internalstandard), MeOH-d₄ (referenced to the residual solvent signal), orDMSO-d₆ (referenced to the residual solvent signal). Chemical shifts aregiven in ppm (δ-scale), coupling constants (J) in Hz. Completeassignment of all NMR signals was performed using a combination ofH,H-COSY, H,H-ROESY, H,C-HSQC and H,C-HMBC experiments. Mass spectrawere measured using FAB (ionization by Xe, accelerating voltage 8 kV,glycerol+thioglycerol matrix) or ESI (electrospray). Chromatographies onreverse phase were performed on a Biotage SP1 apparatus, HPFC systemwith KP-C18-HS, 25+M, 35-70 mm, 90 Å or 40+M, as solid support.

Example 14-Amino-5-phenyl-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine (2a)

An argon purged mixture of 7-iodotubericidine 1 (200 mg, 0.51 mmol) {forpreparation, see Seela, F.; Ming, X. Tetrahedron 2007, 63, 9850-9861},phenylboronic acid (93 mg, 0.76 mmol), Na₂CO₃ (502 mg, 4.74 mmol),Pd(OAc)₂ (6.6 mg, 0.029 mmol) and TPPTS (42 mg, 0.07 mmol) in water/MeCN(2:1, 3.6 ml) was stirred at 80° C. for 1 h. After removal of volatilesin vacuo the residue was purified by reverse phase chromatography(0→100% MeOH in water) affording title compound 2a as white solid (94mg, 54%). Mp 119° C. [α]²⁰ _(D) −49.8 (c 0.301, MeOH). ¹H NMR (600 MHz,DMSO-d₆): 3.53 (ddd, 1H, J_(gem)=11.9, J_(5′b,OH)=6.1, J_(5′b,4′)=3.9,H-5′b); 3.63 (ddd, 1H, J_(gem)=11.9, J_(5′a,OH)=5.2, J_(5′a,4′)=3.4,H-5′a); 3.90 (ddd, 1H, J_(4′,5′)=3.9, 3.4, J_(4′,3′)=3.5, H-4′); 4.10(bm, 1H, H-3′); 4.46 (bm, 1H, H-2′); 5.16 (d, 1H, J_(OH,3′)=3.5, OH-3′);5.22 (dd, 1H, J_(OH,5′)=6.1, 5.2, OH-5′); 5.36 (d, 1H, J_(OH,2′)=4.8,OH-2′); 6.12 (d, 1H, J_(1′,2′)=6.3, H-1′); 7.37 (m, 1H, H-p-Ph); 7.47(m, 2H, H-o-Ph); 7.49 (m, 2H, H-m-Ph); 7.54 (s, 1H, H-6); 8.15 (s, 1H,H-2). ¹³C NMR (151 MHz, DMSO-d₆): 61.93 (CH₂-5′); 70.89 (CH-3′); 74.05(CH-2′); 85.38 (CH-4′); 87.27 (CH-1′); 100.73 (C-4a); 116.57 (C-5);121.41 (CH-6); 127.20 (CH-p-Ph); 128.70 (CH-o-Ph); 129.28 (CH-m-Ph);134.71 (C-i-Ph); 151.10 (C-7a); 151.95 (CH-2); 157.57 (C-4).). IR (KBr):3479, 3391, 1623, 1585, 1566, 1538, 1489, 1466, 1445, 1296, 1216, 1182,1157, 1147, 1119, 1083, 1047, 1028, 1000, 798, 762, 705, 615, 503. MS(FAB) m/z 343 (M+H), 365 (M+Na). HRMS (FAB) for C₁₇H₁₉N₄O₄ [M+H] calcd:343.1406. found: 343.1409. Anal. Calcd for C₁₇H₁₈N₄O₄.0.8H₂O: C, 57.23;H, 5.54; N, 15.70. Found: C, 57.44; H, 5.27; N, 15.43.

Example 24-Amino-5-(4-methoxyphenyl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2b)

Title compound was prepared by following the procedure in Example 1.Pale yellow solid after lyophilization. Yield 36%. Mp 121° C. [α]²⁰ _(D)−21.2 (c 0.304, MeOH). ¹H NMR (600 MHz, DMSO-d₆): 3.53 (ddd, 1H,J_(gem)=11.9, J_(5′b,OH)=6.3, J_(5′b,4)=3.8, H-5′b); 3.62 (ddd, 1H,J_(gem)=11.9, J_(5′a,OH)=4.8, J_(5′a,4′)3.8, H-5′a); 3.80 (s, 3H, CH₃O);3.90 (td, 1H, J_(4′,5′)=3.8, J_(4′,3′)=3.1, H-4′); 4.09 (ddd, 1H,J_(3′,2′)=5.1, J_(3,OH)=4.7, J_(3′,4′)=3.1, H-3′); 4.45 (ddd, 1H,J_(2′,OH)=6.5, J_(2′,1′)=6.3, J_(2′,3′)=5.1, H-2′); 5.14 (d, 1H,J_(OH,3′)=4.7, OH-3′); 5.22 (dd, 1H, J_(OH,5′)=6.3, 4.8, OH-5′); 5.33(d, 1H, J_(OH,2′)=6.5, OH-2′); 6.10 (d, 1H, J_(1′,2′)′=6.3, H-1′); 7.05(m, 2H, H-m-C₆H₄OMe); 7.38 (m, 2H, H-o-C₆H₄OMe); 7.45 (s, 1H, 1H-6);8.13 (s, 1H, H-2). ¹³C NMR (151 MHz, DMSO-d₆): δ5.44 (CH₃O); 61.94(CH₂-5′); 70.90 (CH-3′); 74.01 (CH-2′); 85.34 (CH-4′); 87.24 (CH-1′);100.96 (C-4a); 114.70 (CH-m-C₆H₄OMe); 116.20 (C-5); 120.81 (CH-6);126.85 (C-i-C₆H₄OMe); 129.97 (CH-o-C₆H₄OMe); 150.88 (C-7a); 151.86(CH-2); 157.59 (C-4); 158.68 (C-p-C₆H₄OMe). IR (KBr): 3470, 3391, 2836,1630, 1620, 1586, 1565, 1540, 1506, 1466, 1442, 1421, 1292, 1246, 1216,1175, 1147, 1117, 1109, 1083, 1055, 1030, 838, 796, 706, 637. MS (FAB)m/z 373 (M+H), 395 (M+Na). HRMS (FAB) for C₁₈H₂₁N₄O₅ [M+H] calcd:373.1512. found: 373.1498; for C₁₈H₂₀NaN₄O₅ [M+Na] calcd: 395.1331.found: 395.1327. Anal. Calcd for C₁₈H₂₀N₄O₅.0.95H₂O: C, 55.51; H, 5.67;N, 14.38. Found: C, 55.59; H, 5.44; N, 14.04.

Example 34-Amino-5-[4-(methylthio)phenyl]-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2c)

Title compound was prepared by following the procedure in Example 1.White needles after recrystallization from MeOH. Yield 48%. Mp 227-228°C. [α]²⁰ _(D) −67.3 (c 0.237, DMSO). ¹H NMR (500 MHz, DMSO-d₆): 2.52 (s,3H, CH₃S); 3.53 (ddd, 1H, J_(gem)=12.0, J_(5′b,OH)=6.3, J_(5′,4′)=3.9,H-5′b); 3.63 (ddd, 1H, J_(gem)=12.0, J_(5′a,OH)=5.1, J_(5′a,4′)=3.8,H-5′a); 3.90 (ddd, 1H, J_(4′,5′)=3.9, 3.8, J_(4′,3′)=3.1, H-4′); 4.10(ddd, 1H, J_(3′,2′)=5.1, J_(3,OH)=4.8, J_(3′,4′)=3.1, H-3′); 4.45 (ddd,1H, J_(2′,OH)=6.5, J_(2′,1′)=6.2, J_(2′,3′)=5.1, H-2′); 5.12 (d, 1H,J_(OH,3′)=4.8, OH-3′); 5.20 (dd, 1H, J_(OH,5′)=6.3, 5.1, OH-5′); 5.32(d, 1H, J_(OH,2′)=6.5, OH-2′); 6.11 (d, 1H, J_(1′,2′)=6.2, H-1′); 6.16(bs, 2H, NH₂); 7.37 (m, 2H, H-m-C₆H₄SMe); 7.41 (m, 2H, H-o-C₆H₄SMe);7.52 (s, 1H, H-6); 8.14 (s, 1H, H-2). ¹³C NMR (125.7 MHz, DMSO-d₆):14.98 (CH₃S); 61.88 (CH₂-5′); 70.83 (CH-3′); 73.99 (CH-2′); 85.32(CH-4′); 87.24 (CH-1′); 100.67 (C-4a); 116.01 (C-5); 121.25 (CH-6);126.72 (CH-m-C₆H₄SMe); 129.12 (CH-o-C₆H₄SMe); 131.15 (C-i-C₆H₄SMe);136.90 (C-p-C₆H₄SMe); 151.07 (C-7a); 151.90 (CH-2); 157.55 (C-4). IR(KBr): 3476, 3440, 3343, 3318, 3200, 3123, 2693, 1630, 1584, 1570, 1549,1534, 1492, 1465, 1430, 1402, 1298, 1269, 1212, 1177, 1147, 1124, 1096,1080, 1054, 1016, 967, 832, 796, 716, 690. MS (FAB): m/z 389 (M+H). HRMS(FAB) for C₁₈H₂₁N₄O₄S [M+H] calcd: 389.1284. found: 389.1282. Anal.Calcd for C₁₈H₂₀N₄O₄S: C, 55.66; H, 5.19; N, 14.42. Found: C, 55.28; H,5.25; N, 14.16.

Example 44-Amino-5-(naphtalen-1-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2d)

Title compound was prepared by following the procedure in Example 1.Crude product was prepurified by chromatography on silica (0→20% MeOH inCHCl₃) before final reverse phase chromatography. White solid afterlyophilization. Yield 47%. Mp 134° C. [α]²⁰ _(D) −56.1 (c 0.292, MeOH).¹H NMR (500 MHz, DMSO-d₆, T=353 K): 3.57 (bddd, 1H, J_(gem)=12.0,J_(5′b,OH)=5.1, J_(5′,4′)=3.8, H-5′b); 3.67 (bdt, 1H, J_(gem)=12.0,J_(5′a,OH)=J_(5′a,4′)4.0, H-5′a); 3.96 (ddd, 1H, J_(4′,5′)=4.0, 3.8,J_(4′,3′)=3.1, H-4′); 4.17 (bm, 1H, H-3′); 4.54 (bm, 1H, H-2′); 4.88(bs, 1H, OH-3′); 4.95 (bdd, 1H, J_(OH,5′)=5.1, 4.8, OH-5′); 5.13 (bs,1H, OH-2′); 5.43 (bs, 2H, NH₂); 6.19 (d, 1H, J_(1′,2′)=5.9, H-1′);7.49-7.53 (m, 3H, H-6 and H-2,7-naphth); 7.56 (ddd, 1H, J_(6,5)=8.1,J_(6,7)=6.9, J_(8,7)=1.1, H-6-naphth); 7.61 (dd, 1H, J_(3,4)=8.2,J_(3,2)=7.1, H-3-naphth); 7.81 (bd, 1H, J_(8,7)=8.4, H-8-naphth); 7.99(bd, 1H, J_(4,3)=8.2, H-4-naphth); 8.01 (bd, 1H, J_(5,6)=8.1,H-5-naphth); 8.18 (s, 1H, H-2). ¹³C NMR (151 MHz, DMSO-d₆, T=353 K):61.66 (CH₂-5′); 70.53 (CH-3′); 73.90 (CH-2′); 85.06 (CH-4′); 87.75(CH-1′); 102.65 (C-4a); 113.04 (C-5); 122.03 (CH-6); 125.38(CH-3-naphth); 125.46 (CH-8-naphth); 125.99 (CH-6-naphth); 126.38(CH-7-naphth); 127.82 (CH-4-naphth); 128.05 (CH-2-naphth); 128.10(CH-5-naphth); 131.55 (C-1-naphth); 132.10 (C-8a-naphth); 133.41(C-4a-naphth); 150.43 (C-7a); 151.66 (CH-2); 157.09 (C-4). IR (KBr):3478, 3436, 3392, 3240, 3057, 1632, 1621, 1585, 1569, 1535, 1506, 1469,1398, 1296, 1257, 1108, 1081, 1046, 1016, 946, 849, 805, 797, 790, 779,740. MS (FAB): m/z 393 (M+H). HRMS (FAB) for C₂₁H₂₁N₄O₄ [M+H] calcd:393.1563. found: 393.1564. Anal. Calcd for C₂₁H₂₀N₄O₄.0.8H₂O: C, 62.00;H, 5.35; N, 13.77. Found: C, 62.25; H, 5.21; N, 13.52.

Example 54-Amino-5-(naphtalen-2-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2e)

Title compound was prepared by following the procedure in Example 1.Crude product was prepurified by chromatography on silica (0→10% MeOH inCHCl₃) before final reverse phase chromatography. White solid afterlyophilization. Yield 18%. Mp 129° C. [α]²⁰ _(D) −59.8 (c 0.246, MeOH).¹H NMR (500 MHz, DMSO-d₆): 3.55 (ddd, 1H, J_(gem)=11.7, J_(5′b,OH)=6.2,J_(5′,4′)=3.5, H-5′b); 3.65 (ddd, 1H, J_(gem)=11.7, J_(5′a,OH)=5.1,J_(5′a,4′)=3.5, H-5′a); 3.92 (q, 1H, J_(4′,5′)=J_(4′,3′)=3.5, H-4′);4.13 (ddd, 1H, J_(3′,2′)=5.2, J_(3,OH)=4.7, J_(3′,4′)=3.5, H-3′); 4.49(ddd, 1H, J_(2′,OH)=6.5, J_(2′,1′)=6.2, J_(2′,3′)=5.2, H-2′); 5.15 (d,1H, J_(OH,3′)=4.7, OH-3′); 5.22 (dd, 1H, J_(OH,5′)=6.2, 5.1, OH-5′);5.36 (d, 1H, J_(OH,2′)=6.5, OH-2′); 6.15 (d, 1H, J_(1′,2′)=6.2, H-1′);6.20 (bs, 2H, NH₂); 7.53 (td, 1H, J_(6,5)=J_(6,7)=8.2, J_(6,8)=1.4,H-6-naphth); 7.56 (td, 1H, J_(7,6)=J_(7,8)=8.2, J_(7,6)=1.4,H-7-naphth); 7.65 (dd, 1H, J_(3,4)=8.6, J_(3,1)=1.7, H-3-naphth); 7.66(s, 1H, H-6); 7.97-8.00 (m, 3H, H-1,5,8-naphth); 8.03 (d, 1H,J_(4,3)=8.6, H-4-naphth); 8.17 (s, 1H, H-2). ¹³C NMR (125.7 MHz,DMSO-d₆): δ1.86 (CH₂-5′); 70.77 (CH-3′); 74.00 (CH-2′); 85.29 (CH-4′);87.32 (CH-1′); 100.83 (C-4a); 116.50 (C-5); 121.66 (CH-6); 126.06(CH-6-naphth); 126.70 (CH-7-naphth); 126.79 (CH-1-naphth); 127.13(CH-3-naphth); 127.79 and 127.95 (CH-5,8-naphth); 128.63 (CH-4-naphth);132.00 and 132.10 (C-1,4a-naphth); 133.40 (C-8a-naphth); 151.20 (C-7a);151.90 (CH-2); 157.57 (C-4). IR (KBr): 3475, 3438, 3392, 3240, 3054,1630, 1621, 1584, 1566, 1538, 1505, 1469, 1376, 1295, 1145, 1119, 1085,1047, 1020, 861, 824, 796, 768, 750, 624, 478. MS (FAB): m/z 393 (M+H).HRMS (FAB) for C₂₁H₂₁N₄O₄ [M+H] calcd: 393.1563. found: 393.1571. Anal.Calcd for C₂₁H₂₀N₄O₄.0.65H₂O: C, 62.41; H, 5.31; N, 13.86. Found: C,62.58; H, 5.18; N, 13.64.

Example 64-Amino-5-(furan-2-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2f)

Title compound was prepared by following the procedure in Example 1. Tansolid after lyophilization. Yield 35%. Mp 118° C. [α]²⁰ _(D) −52.5 (c0.287, MeOH). ¹H NMR (500 MHz, DMSO-d₆): 3.55 and 3.65 (2×dd, 2H,J_(gem)=11.9, J_(5′,4′)=3.9, H-5′); 3.91 (td, 1H, J_(4′,5′)=3.9,J_(4′,3′)=3.4, H-4′); 4.11 (dd, 1H, J_(3,2′)=5.2, J_(3′,4′)=3.4, H-3′);4.41 (dd, 1H, J_(2′,1′)=6.1, J_(2′,3′)=5.2, H-2′); 5.00-5.50 (bs, 2H,OH-2′,3′,5′); 6.09 (d, 1H, J_(1′,2′)=6.1, H-1′); 6.61 (dd, 1H,J_(4,3)=3.3, J_(4,5)=1.9, H-4-furyl); 6.67 (dd, 1H, J_(3,4)=3.3,J_(3,5)=0.8, H-3-furyl); 6.88 (bs, 2H, NH₂); 7.78 (dd, 1H, J_(5,4)=1.9,J_(5,3)=0.8, H-5-furyl); 7.83 (s, 1H, H-6); 8.13 (s, 1H, H-2). ¹³C NMR(125.7 MHz, DMSO-d₆): δ1.81 (CH₂-5′); 70.67 (CH-3′); 74.00 (CH-2′);85.33 (CH-4′); 87.27 (CH-1′); 99.55 (C-4a); 105.50 (CH-3-furyl); 106.34(C-5); 112.09 (CH-4-furyl); 120.70 (CH-6); 142.16 (CH-5-furyl); 148.77(C-2-furyl); 151.04 (C-7a); 152.26 (CH-2); 157.45 (C-4). IR (KBr): 3468,3391, 3252, 1631, 1577, 1562, 1532, 1497, 1456, 1299, 1145, 1121, 1083,1049, 1015, 892, 794, 550. MS (FAB): m/z 333 (M+H), 355 (M+Na). HRMS(FAB) for C₁₅H₁₇N₄O₅ [M+H] calcd: 333.1199. found: 333.1202. Anal. Calcdfor C₁₅H₁₆N₄O₅.1.05H₂O: C, 51.30; H, 5.19; N, 15.95. Found: C, 51.64; H,5.01; N, 15.63.

Example 74-Amino-7-(β-D-ribofuranosyl)-5-(thiophen-2-yl)-7H-pyrrolo[2,3-d]pyrimidine(2g)

Title compound was prepared by following the procedure in Example 1.White solid after recrystallization from MeOH. Yield 32%. Mp 188° C.[α]²⁰ _(D) −74.5 (c 0.235, DMSO). ¹H NMR (500 MHz, DMSO-d₆): 3.54 (ddd,1H, J_(gem)=12.0, J_(5′b,OH)=6.2, J_(5′,4′)=3.8, H-5′b); 3.63 (ddd, 1H,J_(gem)=12.0, J_(5′a,OH)=5.0, =3.8, H-5′a); 3.91 (td, 1H, J_(4′,5′)=3.8,J_(4′,3′)=2.9, H-4′); 4.09 (ddd, 1H, J_(3′,2′)=5.0, J_(3,OH)=4.7,J_(3′,4′)=2.9, H-3′); 4.43 (ddd, 1H, J_(2′,OH)=6.4, J_(2,1)=6.3,J_(2′,3′)=5.0, H-2′); 5.13 (d, 1H, J_(OH,3′)=4.7, OH-3′); 5.20 (dd, 1H,J_(OH,5′)=6.2, 5.0, OH-5′); 5.34 (d, 1H, J_(OH,2′)=6.4, OH-2′); 6.10 (d,1H, J_(1′,2′)=6.3, H-1′); 6.32 (bs, 2H, NH₂); 7.15 (dd, 1H, J_(3,4)=3.5,J_(3,5)=1.2, H-3-thienyl); 7.18 (dd, 1H, J_(4,5)=5.1, J_(4,3)=3.5,H-4-thienyl); 7.57 (dd, 1H, J_(5,4)=5.1, J_(5,3)=1.2, H-5-thienyl); 7.62(s, 1H, H-6); 8.15 (s, 1H, H-2). ¹³C NMR (125.7 MHz, DMSO-d₆): δ1.79(CH₂-5′); 70.78 (CH-3′); 74.06 (CH-2′); 85.40 (CH-4′); 87.17 (CH-1′);100.84 (C-4a); 108.65 (C-5); 122.26 (CH-6); 126.06 (CH-5-thienyl);126.58 (CH-3-thienyl); 128.49 (CH-4-thienyl); 135.72 (C-2-thienyl);150.82 (C-7a); 152.22 (CH-2); 157.49 (C-4). IR (KBr): 3509, 3395, 3322,3220, 3102, 1620, 1590, 1576, 1555, 1508, 1460, 1434, 1349, 1300, 1236,1144, 1119, 1102, 1086, 1063, 1042, 1038, 852, 832, 793, 703, 562, 453.MS (FAB): m/z 349 (M+H). HRMS (FAB) for C₁₅H₁₇N₄O₄S [M+H] calcd:349.0971. found: 349.0965. Anal. Calcd for C₁₅H₁₆N₄O₄S: C, 51.71; H,4.63; N, 16.08. Found: C, 51.33; H, 4.48; N, 15.75.

Example 84-Amino-5-(furan-3-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2h)

Title compound was prepared by following the procedure in Example 1.Pale yellow solid after recrystallization from MeOH. Yield 28%. Mp 190°C. [α]²⁰ _(D) −56.9 (c 0.232, DMSO). ¹H NMR (600 MHz, DMSO-d₆): 3.53(ddd, 1H, J_(gem)=12.0, J_(5′b,OH)=6.2, J_(5′,4′)=3.8, H-5′b); 3.62(ddd, 1H, J_(gem)=12.0, J_(5′a,OH)=4.8, J_(5′a,4′)=3.9, H-5′a); 3.89(ddd, 1H, J_(4′,5′)=3.9, 3.8, J_(4,3′)=3.1, H-4′); 4.04 (bddd, 1H,J_(3′,OH)=3.8, J_(3′,2′)=3.6, J_(3′,4′)=3.1, H-3′); 4.42 (bddd, 1H,J_(2′,1′)=6.3, J_(2′,OH)=5.1, J_(2′,3′)=3.6, H-2′); 5.14 (bd, 1H,J_(OH,3′)=3.8, OH-3′); 5.21 (dd, 1H, J_(OH,5′)=6.2, 4.8, OH-5′); 5.33(bd, 1H, J_(OH,2′)=5.1, OH-2′); 6.08 (d, 1H, J_(1′,2′)=6.3, H-1′); 6.27(bs, 2H, NH₂); 6.70 (dd, 1H, J_(4,5)=1.8, J_(4,2)=0.9, H-4-furyl); 7.50(s, 1H, H-6); 7.81 (dd, 1H, J_(5,4)=1.8, J_(5,2)=1.6, H-5-furyl); 7.83(dd, 1H, J_(2,5)=1.6, J_(2,4)=0.9, H-2-furyl); 8.12 (s, 1H, H-2). ¹³CNMR (151 MHz, DMSO-d₆): δ1.92 (CH₂-5′); 70.84 (CH-3′); 73.96 (CH-2′);85.31 (CH-4′); 87.14 (CH-1′); 101.22 (C-4a); 106.44 (C-5); 111.75(CH-4-furyl); 118.77 (C-3-furyl); 121.17 (CH-6); 139.86 (CH-2-furyl);144.41 (CH-5-furyl); 150.92 (C-7a); 151.97 (CH-2); 157.70 (C-4). IR(KBr): 3512, 3394, 3296, 3252, 1623, 1582, 1561, 1504, 1457, 1364, 1306,1286, 1245, 1152, 1121, 1109, 1067, 1038, 1021, 972, 873, 793, 777. MS(FAB): m/z 333 (M+H). HRMS (FAB) for C₁₅H₁₇N₄O₅ [M+H] calcd: 333.1199.found: 333.1204. Anal. Calcd for C₁₅H₁₆N₄O₅: C, 54.21; H, 4.85; N,16.86. Found: C, 53.82; H, 4.85; N, 16.49.

Example 94-Amino-7-(β-D-ribofuranosyl)-5-(thiophen-3-yl)-7H-pyrrolo[2,3-d]pyrimidine(2i)

Title compound was prepared by following the procedure in Example 1.White-grey solid after recrystallization from MeOH. Yield 56%. Mp 197°C. [α]²⁰ _(D) −58.7 (c 0.237, DMSO). ¹H NMR (600 MHz, DMSO-d₆): 3.53(ddd, 1H, J_(gem)=12.0, J_(5′b,OH)=6.1, J_(5′,4′)=3.8, H-5′b); 3.62(ddd, 1H, J_(gem)=12.0, J_(5′a,OH)=4.9, J_(5′a,4′)=3.9, H-5′a); 3.90(ddd, 1H, =3.9, 3.8, J_(4′,3′)=3.0, H-4′); 4.09 (bddd, 1H,J_(3′,2′)=4.2, J_(3,OH)=3.2, J_(3′,4′)=3.0, H-3′); 4.43 (bddd, 1H,J_(2′,1′)=6.3, ==4.2, H-2′); 5.14 (bd, 1H, J_(OH,3′)=3.2, OH-3′); 5.21(dd, 1H, J_(OH,5′)=6.1, 4.9, OH-5′); 5.34 (bd, 1H, J_(OH,2′)=5.1,OH-2′); 6.09 (d, 1H, J_(1′,2′)=6.3, H-1′); 6.21 (bs, 2H, NH₂); 7.27 (dd,1H, J_(4,5)=4.9, J_(4,2)=1.3, H-4-thienyl); 7.52 (dd, 1H, J_(2,5)=2.9,J_(2,4)=1.3, H-2-thienyl); 7.55 (s, 1H, H-6); 7.72 (dd, 1H, J_(5,4)=5.1,J_(5,2)=2.9, H-5-thienyl); 8.13 (s, 1H, H-2). ¹³C NMR (151 MHz,DMSO-d₆): δ1.90 (CH₂-5′); 70.84 (CH-3′); 74.00 (CH-2′); 85.32 (CH-4′);87.17 (CH-1′); 101.00 (C-4a); 111.19 (C-5); 121.29 (CH-6); 122.24(CH-2-thienyl); 127.62 (CH-5-thienyl); 128.71 (CH-4-thienyl); 134.94(C-3-thienyl); 150.79 (C-7a); 151.96 (CH-2); 157.62 (C-4). IR (ICBr):3509, 3396, 3338, 3240, 3106, 1621, 1593, 1576, 1553, 1509, 1460, 1422,1351, 1299, 1214, 1144, 1121, 1101, 1080, 1061, 1036, 856, 796, 775,713, 562, 456. MS (FAB): m/z 349 (M+H). HRMS (FAB) for C₁₅H₁₇N₄O₄S [M+H]calcd: 349.0971. found: 349.0962. Anal. Calcd for C₁₅H₁₆N₄O₄S: C, 51.71;H, 4.63; N, 16.08. Found: C, 51.51; 1-1, 4.63; N, 15.76.

Example 104-Amino-5-(benzofuran-2-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2j)

Title compound was prepared by following the procedure in Example 1.White needles after recrystallization from MeOH/water. Yield 27%. Mp177° C. [α]²⁰ _(D) −68.8 (c 0.257, MeOH). ¹H NMR (500 MHz, DMSO-d₆):3.57 (ddd, 1H, J_(gem)=12.0, J_(5′b,OH)=6.1, J_(5′,4′)=3.9, H-5′b); 3.67(ddd, 1H, J_(gem)=12.0, J_(5′a,OH)=5.1, J_(5′a,4′)=3.9, H-5′a); 3.93(td, 1H, J_(4′,5′)=3.9, J_(4′,3′)=3.3, H-4′); 4.13 (ddd, 1H,J_(3′,2′)=5.1, J_(3,OH)=4.9, J_(3′,4′)=3.3, H-3); 4.46 (ddd, 1H,J_(2′,OH)=6.3, J_(2′,1′)=6.1, J_(2′,3′)=5.1, H-2); 5.17 (d, 1H,J_(OH,3′)=4.9, OH-3′); 5.23 (dd, 1H, J_(OH,5′)=6.1, 5.1, OH-5′); 5.40(d, 1H, J_(OH,2′)=6.3, OH-2′); 6.14 (d, 1H, J_(1′,2′)=6.1, H-1′); 7.00(bs, 2H, NH₂); 7.13 (d, 1H, J_(3,7)=1.0, H-3-benzofuryl); 7.28 (td, 1H,J_(5,4)=J_(5,6)=7.3, J_(5,7)=1.5, H-5-benzofuryl); 7.30 (td, 1H,J_(6,5)=J_(6,7)=7.3, J_(6,4)=1.7, H-6-benzofuryl); 7.62-7.69 (m, 2H,H-4,7-benzofuryl); 8.11 (s, 1H, H-6); 8.18 (s, 1H, H-2). ¹³C NMR (125.7MHz, DMSO-d₆): δ1.79 (CH₂-5′); 70.70 (CH-3′); 74.12 (CH-2′); 85.46(CH-4′); 87.35 (CH-1′); 99.62 (C-4a); 101.83 (CH-3-benzofuryl); 105.72(C-5); 111.30 (CH-7-benzofuryl); 120.84 (CH-4-benzofuryl); 122.94(CH-6); 123.69 (CH-5-benzofuryl); 124.07 (CH-6-benzofuryl); 129.02(C-3a-benzofuryl); 151.29 (C-2-benzofuryl); 151.35 (C-7a); 152.52(CH-2); 153.99 (C-7a-benzofuryl); 157.52 (C-4). IR (KBr): 3498, 3473,3462, 3379, 3328, 3234, 3197, 3118, 2700, 1639, 1628, 1614, 1576, 1558,1525, 1483, 1474, 1456, 1303, 1262, 1186, 1145, 1127, 1108, 1085, 1056,1010, 884, 806, 792, 785, 746. MS (ESI): m/z 383 (M+H). HRMS (ESI) forC₁₉H₁₉N₄O₅ [M+H] calcd: 383.1350; found: 383.1348. Anal. Calcd forC₁₉H₁₈N₄O₅.0.6H₂O: C, 58.04; H, 4.92; N, 14.25. Found: C, 57.78; H,4.56; N, 14.16.

Example 114-Amino-5-(1H-pyrrol-2-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2k)

An argon purged mixture of 7-iodotubericidine 1 (196 mg, 0.50 mmol),1-N-(Boc)-pyrrole-2-boronic acid (126 mg, 0.60 mmol), Na₂CO₃ (160 mg,1.5 mmol), Pd(OAc)₂ (5.6 mg, 0.025 mmol) and TPPTS (36 mg, 0.06 mmol) inwater/MeCN (2:1, 3 ml) was stirred at 100° C. for 3 h. After cooling themixture was neutralized by the addition of aq HCl (1M), volatiles wereremoved in vacuo and the residue was purified by reverse phasechromatography (0→100% MeOH in water) affording title compound 2k asgreenish solid (127 mg, 77%). Compound was recrystallized from MeOH aswhite solid, after decolorization with active carbon. Mp 205-207° C.[α]_(D) −80.5 (c 0.205, DMSO). ¹H NMR (500.0 MHz, DMSO-d₆): 3.54 (ddd,1H, J_(gem)=11.9, J_(5′b,OH)=6.2, J_(5′b,4′)=3.9, H-5′b); 3.62 (dd, 1H,J_(gem)=11.9, J_(5′a,OH)=5.0, J_(5′a,4′)=3.9, H-5′a); 3.90 (td, 1H,J_(4′,5′)=3.9, J_(4′,3′)=3.1, H-4′); 4.09 (ddd, 1H, J_(3′,2′)=5.1, =4.8,J_(3′,4′)=3.1, H-3′); 4.41 (ddd, 1H, J_(2′,OH)=6.5, J_(2′,1′)=6.3, =5.1,H-2′); 5.17 (d, 1H, J_(OH,3′)=4.8, OH-3′); 5.20 (dd, 1H, J_(OH,5′)=6.2,5.0, OH-5′); 5.33 (d, 1H, J_(OH,2′)=6.5, OH-5′); 6.09 (d, 1H,J_(1′,2′)=6.3, H-1′); 6.13 (ddd, 1H, J_(3,4)=3.3, J_(3,NH)=2.4,J_(3,5)=1.5, H-3-pyrr); 6.16 (ddd, 1H, J_(4,3)=3.3, J_(4,5)=2.7,J_(4,NH)=2.4, H-4-pyrr); 6.32 (bs, 2H, NH₂); 6.85 (td, 1H,J_(5,4)=J_(5,NH)=2.7, J_(5,3)=1.5, H-5-pyrr); 7.43 (s, 1H, 8.12 (s, 1H,H-2); 11.14 (bs, 1H, NH-pyrr). ¹³C NMR (125.7 MHz, DMSO-d₆): δ2.00(CH₂-5′); 70.92 (CH-3); 74.11 (CH-2); 85.30 (CH-4′); 87.20 (CH-1′);101.09 (C-4a); 107.34 (CH-3-pyrr); 108.68 (C-5); 109.06 (CH-4-pyrr);118.77 (CH-5-pyrr); 120.55 (CH-6); 124.69 (C-2-pyrr); 150.35 (C-7a);151.98 (CH-2); 157.62 (C-4). MS (ESI) m/z 332 (M+H), 354 (M+Na). HRMS(ESI) for C₁₅H₁₈N₅O₄ [M+H] calcd: 332.1353. found: 332.1354. Anal. Calcdfor C₁₅H₁₇N₅O₄.⅓H₂O: C, 53.41; H, 5.28; N, 20.76. Found: C, 53.32; H,5.16; N, 20.57.

Example 124-Amino-5-(1H-pyrrol-3-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2l)

An argon purged mixture of 7-iodotubericidine 1 (893 mg, 2.28 mmol),1-(triisopropylsilyl)-1H-pyrrole-3-boronic acid (822 mg, 3.08 mmol),Na₂CO₃ (724 mg, 6.83 mmol), Pd(OAc)₂ (26 mg, 0.12 mmol) and TPPTS (162mg, 0.28 mmol) in water/MeCN (2:1, 18 ml) was stirred at 100° C. for 18h. After cooling the mixture was neutralized by the addition of aq HCl(1M) and desalted by reverse phase chromatography (0→100% MeOH in water)affording crude product contaminated by starting iodide. Re-purificationby column chromatography on silica (6% MeOH in CHCl₃) afforded titlecompound 21 as white solid (480 mg, 63%). Compound was recrystallizedfrom EtOH. Mp 188-190° C. [α]_(D) −59.8 (c 0.276, DMSO). ¹H NMR (500.0MHz, DMSO-d₆): 3.52 (ddd, 1H, J_(gem)=12.0, J_(5′b,OH)=6.3,J_(5′b,4′)=3.8, H-5′b); 3.61 (dd, 1H, J_(gem)=12.0, J_(5′a,OH)=4.9,J_(5′a,4′)3.8, H-5′a); 3.88 (td, 1H, J_(4′,5′)=3.8, J_(4′,3′)=3.0,H-4′); 4.08 (ddd, 1H, J_(3′,2′)=5.1, J_(3′,OH)=4.7, J_(3′,4′)=3.0,H-3′); 4.43 (ddd, 1H, J_(2′,OH)=6.5, J_(2′,1′)=6.4, J_(2′,3′)=5.1,H-2′); 5.11 (d, 1H, J_(OH,3′)=4.7, OH-3′); 5.24 (dd, 1H, J_(OH,5′)=6.3,4.9, OH-5′); 5.30 (d, 1H, J_(OH,2′)=6.5, OH-5′); 6.06 (d, 1H, =6.4,H-1′); 6.18 (td, 1H, J_(4,5)=J_(4,NH)=2.6, J_(4,2)=1.6, H-4-pyrr); 6.26(bs, 2H, NH₂); 6.88 (dt, 1H, J_(2,NH)=2.6, J_(2,4)=J_(2,5)=1.6,H-2-pyrr); 6.90 (td, 1H, J_(5,4)=J_(5,NH)=2.6, J_(5,2)=1.6, H-5-pyrr);7.27 (s, 1H, H-6); 8.08 (s, 1H, H-2); 11.03 (bs, 1H, NH-pyrr). ¹³C NMR(125.7 MHz, DMSO-d₆): δ2.00 (CH₂-5′); 70.94 (CH-3′); 73.89 (CH-2′);85.23 (CH-4′); 87.12 (CH-1′); 101.65 (C-4a); 108.29 (CH-4-pyrr); 111.17(C-5); 115.87 (C-3-pyrr); 116.44 (CH-2-pyrr); 119.33 (CH-2-pyrr); 119.65(CH-6); 150.42 (C-7a); 151.62 (CH-2); 157.77 (C-4). MS (ESI) m/z 332(M+H), 354 (M+Na). HRMS (ESI) for C₁₅H₁₈N₅O₄ [M+H] calcd: 332.1353.found: 332.1354. Anal. Calcd for C₁₅H₁₇N₅O₄.0.8H₂O: C, 52.11; H, 5.42;N, 20.26. Found: C, 52.32; H, 5.32; N, 20.03.

Example 134-Amino-5-(1H-pyrazol-4-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2m)

Method A. Title compound was prepared by following the procedure inExample 1. White solid. Yield 9%. Mp 156° C. ¹H NMR (500 MHz,DMSO-d₆+DCl): 3.53 and 3.61 (2×dd, 2×1H, J_(gem)=11.9, J_(5′,4′)=3.8,H-5′); 3.93 (td, 1H, J_(4′,5′)=3.8, J_(4′,3′)=3.3, H-4′); 4.10 (dd, 1H,J_(3′,2′)=4.9, J_(3′,4′)=3.3, H-3′); 4.35 (dd, 1H, J_(2′,1′=)6.1,J_(2′,3′)=4.9, H-2′); 6.15 (d, 1H, J_(1′,2′)=6.1, H-1′); 7.85 (s, 1H,H-6); 8.09 (s, 2H, H-3,5-pyrazole); 8.52 (s, 1H, H-2). ¹³C NMR (125.7MHz, DMSO-d₆+DCl): 61.52 (CH₂-5′); 70.72 (CH-3′); 74.83 (CH-2′); 85.97(CH-4′); 87.14 (CH-1′); 99.99 (C-4a); 109.14 (C-4-pyrazole); 112.19(C-5); 124.02 (CH-6); 133.06 (CH-3,5-pyrazole); 142.88 (CH-2); 148.10(C-7a); 151.41 (C-4). IR (CH-1′): 3468, 3338, 3239, 3200, 3115, 1740,1629, 1583, 1559, 1523, 1469, 1306, 1123, 1076, 1044, 1024, 796. MS(ESI): m/z 333 (M+H), 355 (M+Na). HRMS (ESI) for C₁₄H₁₇N₆O₄ [M+H] calcd:333.1306. found: 333.1304.Method B. An argon purged mixture of 7-iodotubericidine 1 (1.649 g, 4.2mmol), 1-dimethylsulfamoyl-4-tributylstannylpyrazole {for preparation,see US 20040157892 A1} (2.97 g, 6.4 mmol), PdCl₂(PPh₃)₂ (148 mg, 0.21mmol) in DMF (15 ml) was stirred at 100° C. for 3 h. Volatiles wereremoved under reduced pressure, the residue was several timesco-evaporated with toluene/MeOH and finally with silica. Columnchromatography on silica (0→8% MeOH in CHCl₃) afforded product protectedon pyrazole nitrogen by dimethylsulfamoyl group (1.459 g, 79%). Thismaterial was directly deprotected by addition of aq HCl (1M, 20 ml) andstirring at 100° C. for 3 h. Volatiles were removed in vacuo, and theresidue was co-evaporated with water (10-15×), once with aq ammonia (25%w/w) and again with water (6×). Re-crystallization from water affordedtitle compound 2m as white needles (707 mg, 64% yield of deprotectionstep). Mother liquors were purified by reverse phase chromatography(0→100% MeOH in water) affording after crystallization from MeOH/wateradditional portion of title compound (243 mg, 22% yield of deprotectionstep). Overall yield (coupling+deprotection) is 68%.

Example 144-Amino-5-(1H-pyrazol-3-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2n)

An argon purged mixture of 7-iodotubericidine 1 (392 mg, 1 mmol),pyrazole-5-boronic acid (224 mg, 2 mmol), Na₂CO₃ (318 mg, 3 mmol),Pd(OAc)₂ (11 mg, 0.05 mmol) and TPPTS (71 mg, 0.125 mmol) in water/MeCN(2:1, 5 ml) was stirred at 100° C. for 18 h. After cooling the mixturewas neutralized by the addition of aq HCl (1M) and desalted by reversephase chromatography (0→100% MeOH in water) and re-purified by columnchromatography on silica (8% MeOH in CHCl₃) affording title compound 2nas white glassy solid (273 mg, 82%). White solid after recrystallizationfrom boiling water. Mp 135° C. ¹H NMR (500 MHz, DMSO-d₆): 3.55 (ddd, 1H,J_(gem)=12.1, J_(5′b,OH)=6.4, J_(5′b,4′)=4.1, H-5′b); 3.66 (ddd, 1H,J_(gem)=12.1, J_(5′a,OH)=5.0, J_(5′a,4′)=3.9, H-5′a); 3.90 (ddd, 1H,J_(4′,5′)=4.1, 3.9, J_(4′,3′)=3.2, H-4′); 4.11 (ddd, 1H, J_(3,2′)=5.2,J_(3′,2′)=4.8, J_(3′,4)=3.2, H-3′); 4.44 (ddd, 1H, J_(2′,OH)=6.4,J_(2′,1′)=6.3, J_(2′,3′)=5.2, H-2′); 6.04 (d, 1H, J_(1′,2′)=6.1, H-1′);6.66 (dd, 1H, J_(4,5)=2.4, J_(4,NH)=1.9, H-4-pyrazole); 7.25 (bs, 1H,NH_(a)H_(b)); 7.81 (dd, 1H, J_(5,4)=2.4, J_(5,NH)=1.5, H-5-pyrazole);7.86 (s, 1H, H-6); 8.04 (s, 1H, H-2); 9.24 (bs, 1H, NH_(a)H_(b)); 12.88(bs, 1H, NH). ¹³C NMR (125.7 MHz, DMSO-d₆): δ1.98 (CH₂-5′); 70.76(CH-3′); 73.75 (CH-2′); 85.27 (CH-4′); 87.32 (CH-1′); 100.36 (C-4a);101.91 (CH-4-pyrazole); 109.76 (C-5); 120.64 (CH-6); 130.20(CH-5-pyrazole); 146.26 (C-3-pyrazole); 151.02 (C-7a); 152.44 (CH-2);158.47 (C-4). IR (KBr): 3411, 3290, 3136, 2665, 1633, 1597, 1576, 1550,1474, 1301, 1138, 1109, 1082, 1050, 1019, 934, 798, 765, 651. MS (ESI):m/z 333 (M+H), 355 (M+Na). HRMS (ESI) for C₁₄H₁₇N₆O₄ [M+H] calcd:333.1306. found: 333.1306. Anal. Calcd for C₁₄H₁₆N₆O₄.1.85H₂O: C, 45.99;H, 5.43; N, 22.98. Found: C, 46.22; H, 5.44; N, 22.68.

Example 154-Amino-5-ethynyl-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine (2o)

Step 1.4-Amino-7-(β-D-ribofuranosyl)-5-[(trimethylsilyl)ethynyl]-7H-pyrrolo[2,3-d]pyrimidine(3)

An argon purged mixture of 7-iodotubericidine 1 (1010 mg, 2.57 mmol),PdCl₂(PPh₃)₂ (90 mg, 0.13 mmol), CuI (49 mg, 0.26 mmol),trimethylsilylacetylene (3.6 ml, 25.7 mmol) and triethylamine (1 ml) wasstirred in DMF (4 ml) at RT for 16 h. Volatiles were removed in vacuoand the rest was twice co-evaporated with EtOH and loaded on silica byco-evaporation from EtOH. Column chromatography on silica (0→3% MeOH inCHCl₃) afforded product 3 as off-white crystalline solid (962 mg,quantitative). Product was recrystallized from CHCl₃MeOH. Mp 167-169° C.[α]_(D) −77.6 (c 0.313, DMSO). ¹H NMR (400.0 MHz, DMSO-d₆): 0.24 (s, 9H,CH₃-TMS); 3.54 (ddd, 1H, J_(gem)=12.0, J_(5′b,OH)=6.2, J_(5′b,4′)=3.8,H-5′b); 3.63 (dd, 1H, J_(gem)=12.0, J_(5′a,OH)=5.0, J_(5′a,4′)=3.8,H-5′a); 3.90 (td, 1H, J_(4′,5′)=3.8, J_(4′,3′)=3.2, H-4′); 4.08 (ddd,1H, J_(3′,2′)=5.0, J_(3′,OH)=4.8, J_(3′,4′)=3.2, H-3′); 4.36 (ddd, 1H,J_(2′,OH)=6.3, =6.0, J_(2′,3′)=5.0, H-2′); 5.15 (d, 1H, J_(OH,3′)=4.7,OH-3′); 5.20 (dd, 1H, J_(OH,5′)=6.2, 5.0, OH-5′); 5.35 (d, 1H,J_(OH,2′)=6.3, OH-5′); 6.02 (d, 1H, J_(1′,2′)=6.0, H-1′); 6.64 (bs, 2H,NH₂); 7.84 (s, 1H, H-6); 8.13 (s, 1H, H-2). ¹³C NMR (100.6 MHz,DMSO-d₆): −0.19 (CH₃-TMS); 61.47 (CH₂-5′); 70.45 (CH-3′); 74.06 (CH-2′);85.26 (CH-4′); 87.23 (CH-1′); 94.60 (C-5); 96.78 (—C≡CTMS); 99.14(—C≡CTMS); 102.41 (C-4a); 127.38 (CH-6); 149.48 (C-7a); 152.82 (CH-2);157.58 (C-4). MS (ESI) m/z 363 (M+H), 385 (M+Na). HRMS (ESI) forC₁₆H₂₃N₄O₄Si [M+H] calcd: 363.1483. found: 363.1484. Anal. Calcd forC₁₆H₂₂N₄O₄Si: C, 53.02; H, 6.12; N, 15.46. Found: C, 52.91; H, 6.11; N,15.30.

Step 2.4-Amino-5-ethynyl-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine (2o)

A mixture of the compound 3 from Step 1 (686 mg, 1.89 mmol) and K₂CO₃(130 mg, 0.94 mmol) in MeOH (15 ml) was stirred at RT for 1 h, followedby the co-evaporation with silica. Column chromatography on silica (5%MeOH in CHCl₃) provided title compound 2o as white crystalline solid(528 mg, 96%). Compound was recrystallized from MeOH/water as long ochryneedles. Mp 215-217° C. [α]_(D) −88.3 (c 0.524, DMSO). ¹H NMR (500.0MHz, DMSO-d₆): 3.53 (ddd, 1H, J_(gem)=12.0, J_(5′b,OH)=6.2,J_(5′b,4′)=3.8, H-5′b); 3.63 (dd, 1H, J_(gem)=12.0, J_(5′a,OH)=5.0,J_(5′a,4′)=3.8, H-5′a); 3.89 (td, 1H, J_(4′,5′)=3.8, J_(4′,3′)=3.2,H-4′); 4.07 (ddd, 1H, J_(3′,2′)=5.0, J_(3,OH)=4.8, J_(3′,4′)=3.2, H-3′);4.29 (s, 1H, HC≡C—); 4.37 (ddd, 1H, J_(2′,OH)=6.3, J_(2′,1′)6.1,J_(2′,3′)5.0, H-2′); 5.16 (d, 1H, J_(OH,3′)=4.8, OH-3′); 5.23 (dd, 1H,J_(OH,5′)=6.2, J_(2′,3′)5.0, OH-5′); 5.38 (d, 1H, J_(OH,2′)=6.3, OH-5′);6.01 (d, 1H, J_(1′,2′)=6.1, H-1′); 6.70 (bs, 2H, NH₂); 7.83 (s, 1H,H-6); 8.12 (s, 1H, H-2). ¹³C NMR (125.7 MHz, DMSO-d₆): δ1.73 (CH₂-5′);70.74 (CH-3′); 74.21 (CH-2′); 77.52 (—CCH); 83.37 (—CCH); 85.51 (CH-4′);87.42 (CH-1′); 94.17 (C-5); 102.65 (C-4a); 127.74 (CH-6); 149.73 (C-7a);153.04 (CH-2); 157.78 (C-4). MS (ESI) m/z 291 (M+H), 313 (M+Na). HRMS(ESI) for C₁₃H₁₅N₄O₄ [M+H] calcd: 291.1088. found: 291.1088. Anal. Calcdfor C₁₃H₁₄N₄O₄: C, 53.79; H, 4.86; N, 18.91. Found: C, 53.34; H, 4.95;N, 18.97.

Example 164-Amino-7-(β-D-ribofuranosyl)-5-(1H-1,2,3-triazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine(2p)

An argon purged mixture of 7-ethynyltubericidine 2o {compound fromexample 15} (200 mg, 0.69 mmol), CuI (6.5 mg, 0.03 mmol) and TMSN₃ (150μl, 1.15 mmol) in MeOH (0.2 ml)/DMF (1.8 ml) was stirred 100° C. for 24h. Volatiles were evaporated under reduced pressure, the residue wastwice co-evaporated with MeOH and rest was suspended in MeOH andfiltered through celite. Filtrate was co-evaporated with silica andcolumn chromatography on silica afforded title compound 2p as yellowishsolid (55 mg, 24%). Compound was recrystallized from water. Mp 286-288°C. [α]_(D) −70 (c 0.217, DMSO). ¹H NMR (499.8 MHz, DMSO-d₆, t=60° C.):3.58 (dd, 1H, J_(gem)=11.9, J_(5′b,4′)=4.2, H-5′b); 3.68 (dd, 1H,J_(gem) 11.9, J_(5′a,4′)=3.9, H-5′a); 3.93 (ddd, 1H, J_(4′,5′)=4.2, 3.9,J_(4′,3′)=3.6, H-4′); 4.15 (dd, 1H, J_(3′,2′)=5.3, J_(3′,4′)=3.6, H-3′);4.45 (dd, 1H, J_(2′,1′)=6.0, J_(2′,3′)=5.3, H-2′); 4.93, 5.04 and 5.17(3×bs, 3×1H, OH-2′,3′,5′); 6.08 (d, 1H, J_(1′,2′)=6.0, H-1′); 7.85 (bs,2H, NH₂); 7.91 (s, 1H, H-6); 8.09 (s, 1H, H-2); 8.23 (s, 1H,H-5-triazole). ¹³C NMR (125.7 MHz, DMSO-d₆, t=60° C.): 61.83 (CH₂-5′);70.54 (CH-3′); 73.72 (CH-2′); 85.15 (CH-4′); 87.41 (CH-1′); 100.16(C-4a); 105.83 (C-5); 120.80 (CH-6); 126.16 (CH-5-triazole); 141.51(C-4-triazole); 151.04 (C-7a); 152.35 (CH-2); 158.04 (C-4). MS (ESI) m/z334 (M+H), 356 (M+Na). HRMS (ESI) for C₁₃H₁₆N₇O₄ [M+H] calcd: 334.1258.found: 334.1258. Anal. Calcd for C₁₃H₁₅N₇O₄: C, 46.85; H, 4.54; N,29.42. Found: C, 47.12; H, 4.82; N, 27.96.

Example 174-Amino-7-(β-D-ribofuranosyl)-5-(thiazol-2-yl)-7H-pyrrolo[2,3-d]pyrimidine(2q)

Step 1.4-Amino-5-iodo-7-[2,3,5-tris-O-(tert-butyldimethylsilyl)-β-D-ribofuranosyl]-7H-pyrrolo[2,3-d]pyrimidine(4)

tert-Butyldimethylsilyl chloride (6.78 g, 45 mmol) was added to asolution of 7-iodotubericidine 1 (3.92 g, 10 mmol) and imidazole (6.12g, 90 mmol) in DMF (20 ml) and the mixture was stirred overnight at RT.The mixture was diluted with hexane (100 ml) and washed with aq NaCl(5%, 100 ml). Aqueous layer was re-extracted with hexane (2×25 ml).Organic phase was washed with aq NaCl (5% w/w, 4×50 ml). Collectedorganic extracts were dried over MgSO₄, evaporated and chromatographedon silica (hexanes/AcOEt, 50:1→5:1) affording title compound 4 ascolorless foam (3.05 g, 41%) and faster moving byproduct resulting from6-amino group N-silylation. This oversilylation byproduct was convertedto title compound 4 by standing of its methanolic solution at RT forseveral days (2.96 g, 40%). Total yield of title compound 4 is 81%. ¹HNMR (500.0 MHz, CDCl₃): −0.28, −0.08, 0.096, 0.098, 0.17 and 0.18 (6×s,6×3H, CH₃Si); 0.77, 0.93 and 0.99 (3×s, 3×9H, (CH₃)C); 3.77 (dd, 1H,J_(gem)=11.4, J_(5′b,4′)=2.3, H-5′b); 3.95 (dd, 1H, J_(gem)=11.4,J_(5′a,4′)=3.0, H-5′a); 4.08 (td, 1H, J_(4′,5′)=3.0, 2.3, J_(4′,3′)=3.0,H-4′); 4.22 (dd, 1H, J_(3′,2′)=4.5, =3.0, H-3′); 4.39 (dd, 1H,J_(2′,1′)=5.6, =4.5, H-2′); 5.70 (bs, 2H, NH₂); 6.26 (d, 1H, =5.6,H-1′); 7.52 (s, 1H, H-6); 8.26 (s, 1H, H-2). ¹³C NMR (125.7 MHz, CDCl₃):−5.33, −5.31, −5.26, −4.76, −4.75 and −4.41 (CH₃Si); 17.84, 18.10 and18.60 (C(CH₃)₃); 25.65, 25.84 and 26.17 ((CH₃)₃C); 50.20 (C-5); 62.94(CH₂-5′); 72.43 (CH-3′); 76.78 (CH-2′); 85.46 (CH-4′); 87.48 (CH-1′);104.26 (C-4a); 126.62 (CH-6); 150.66 (C-7a); 152.19 (CH-2); 156.74(C-4). MS (ESI) m/z 735 (M+H), 757 (M+Na).

Step 2.4-Amino-5-(thiazol-2-yl)-7-[2,3,5-tris-O-(tert-butyldimethylsilyl)-β-D-ribofuranosyl]-7H-pyrrolo[2,3-d]pyrimidine(5)

An argon purged mixture of the compound 4 from Step 1 (544 mg, 0.74mmol), 2-(tributylstannyl)thiazole (554 mg, 1.48 mmol) and PdCl₂(PPh₃)₂(22 mg, 0.03 mmol) in DMF (3 ml) was stirred at 100° C. for 48 h.Volatiles were evaporated under diminished pressure, the residue wasco-evaporated with water (3×), twice with MeOH, twice with hexane andthen co-evaporated with silica from hexane. Column chromatography onsilica (hexanes then hexanes/AcOEt, 10:1→6:1) afforded title compound 5as foam (454 mg, 89%). ¹H NMR (500.0 MHz, CDCl₃): −0.36, −0.10, 0.11,0.13, 0.17 and 0.18 (6×s, 6×3H, CH₃Si); 0.74, 0.95 and 0.99 (3×s, 3×9H,(CH₃)C); 3.79 (dd, 1H, J_(gem)=11.4, J_(5′b,4′)=2.7, H-5′b); 3.96 (dd,1H, J_(gem)=11.4, J_(5′a,4′)=3.2, H-5′a); 4.11 (ddd, 1H, J_(4′,5′)=3.9,2.7, J_(4′,3′)=1.8, H-4′); 4.23 (dd, 1H, =4.7, J_(3′,4′)=1.8, H-3′);4.55 (dd, 1H, J_(2′,1′)=6.9, J_(2′,3′)=4.7, H-2′); 5.96 (bs, 1H,NH_(a)H_(b)); 6.31 (d, 1H, J_(1′,2′)=6.9, H-1′); 7.21 (d, 1H,J_(5,4)=3.4, H-5-thiazolyl); 7.72 (d, 1H, J_(4,5)=3.4, H-4-thiazolyl);7.74 (s, 1H, H-6); 8.26 (s, 1H, H-2); 9.79 (bs, 1H, NH_(a)H_(b)). ¹³CNMR (125.7 MHz, CDCl₃): −5.35, −5.33, −5.19, −4.65, −4.61 and −4.47(CH₃Si); 17.82, 18.13 and 18.57 (C(CH₃)₃); 25.61, 25.86 and 26.15((CH₃)₃C); 63.37 (CH₂-5′); 73.03 (CH-3′); 76.26 (CH-2′); 86.28 (CH-4′);87.01 (CH-1′); 100.52 (C-4a); 112.19 (C-5); 116.76 (CH-5-thiazolyl);122.24 (CH-6); 141.71 (CH-4-thiazolyl); 151.57 (C-7a); 151.94 (CH-2);157.52 (C-4); 163.53 (C-2-thiazolyl). MS (ESI) m/z 692 (M+H), 714(M+Na). HRMS (ESI) for C₃₂H₅₈N₅O₄SSi₃ [M+H] calcd: 692.3512. found:692.3512.

Step 3.4-Amino-7-(β-D-ribofuranosyl)-5-(thiazol-2-yl)-7H-pyrrolo[2,3-d]pyrimidine(2q)

Compound 5 from Step 2 (448 mg, 0.65 mmol) was treated with aq HCl (1M,1 ml) in MeOH (1 ml) at RT for 2 h. Volatiles were removed in vacuo andsolid residue was co-evaporated with water (6×). Crude product wasrecrystallized from MeOH/water affording title compound 2q as white longneedles (132 mg, 58%). Mother liquors were purified by columnchromatography on silica (8% MeOH in CHCl₃) affording additional portionof product (61 mg, 27%). Overall yield 85%. Mp 226-228° C. [α]_(D) −81.5(c 0.254, DMSO). ¹H NMR (600.1 MHz, DMSO-d₆): 3.61 (dd, 1H,J_(gem)=12.0, J_(5′b,4′)=3.6, H-5′b); 3.72 (dd, 1H, J_(gem) 12.0,J_(5′a,4′)=3.9, H-5′a); 3.97 (td, 1H, J_(4′,5′)=3.9, 3.6, J_(4′,3′)=3.9,H-4′); 4.16 (dd, 1H, J_(3′,2′)=4.9, =3.9, H-3′); 4.41 (dd, 1H,J_(2′,1′)=5.6, J_(2,3)=4.9, H-2′); 6.14 (d, 1H, =5.6, H-1′); 7.80 (d,1H, J_(5,4)=3.3, H-5-thiazolyl); 7.91 (d, 1H, J_(4′,5′)=3.3,H-4-thiazolyl); 8.48 (s, 1H, H-2); 8.62 (s, 1H, H-6); 8.99 and 10.88(2×bs, 2H, NH₂). ¹³C NMR (150.9 MHz, DMSO-d₆): δ1.12 (CH₂-5′); 70.19(CH-3′); 74.78 (CH-2′); 85.80 (CH-4′); 87.63 (CH-1′); 99.39 (C-4a);113.09 (C-5); 120.13 (CH-5-thiazolyl); 125.34 (CH-6); 142.09(CH-4-thiazolyl); 145.00 (CH-2); 148.55 (C-7a); 152.47 (C-4); 162.07(C-2-thiazolyl). MS (ESI) m/z 350 (M+H), 372 (M+Na). HRMS (ESI) forC₁₄H₁₆N₅O₄S [M+H] calcd: 350.0918. found: 350.0918. Anal. Calcd forC₁₄H₁₅N₅O₄S.½H₂O.CH₃OH: C, 46.15; H, 5.16; N, 17.94. Found: C, 46.41; H,4.96; N, 17.67.

Example 184-Amino-5-(1H-imidazol-4-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2r)

To a solution of 4-iodo-1-trityl-1H-imidazole (609 mg, 1.4 mmol) in dryTHF (6 ml) was added EtMgBr (1M in THF, 1.5 ml, 1.5 mmol) and themixture was stirred at RT for 10 min. Then a solution of ZnCl₂ (1M inTHF, 2.8 ml, 2.8 mmol) was dropwise added and the mixture was stirredfor 2 h. Resulting thick slurry was transferred to an argon purgedmixture of 2′,3′,5′-tri-O-TBS-7-iodotubericidine {compound 4 fromExample 17, Step 1} (515 mg, 0.7 mmol) and Pd(PPh₃)₄ (40 mg, 0.035 mmol)and the mixture was stirred at 90° C. for 16 h. The mixture was dilutedwith CHCl₃ (20 ml) and washed with aq EDTA (sat., 20 ml). Aqueous layerwas re-extracted with CHCl₃ (2×5 ml). Collected organic extracts weredried over MgSO₄, evaporated and chromatographed on silica(hexanes/AcOEt, 4:1→3:1) affording crude TBS,Tr-protected productcontaminated by N-tritylimidazole. Crude product was deprotected bystirring in aq TFA (90% v/v, 2 ml) at RT for 18 h. The volatiles wereremoved in vacuo and the residue was several times co-evaporated withMeOH and finally loaded on silica by co-evaporation from MeOH. Columnchromatography on silica (0→10% MeOH in CHCl₃) afforded title compound2r as white powder (179 mg, 77%). Compound was recrystallized fromMeOH/water. Mp 276-278° C. [α]_(D) −61.4 (c 0.249, DMSO). ¹H NMR (500.0MHz, DMSO-d₆): 3.53 (ddd, 1H, J_(gem)=12.0, J_(5′b,OH)=6.5,J_(5′b,4′)=4.2, H-5′b); 3.64 (dd, 1H, J_(gem)=12.0, J_(5′a,OH)=4.8,J_(5′a,4′)=4.2, H-5′a); 3.89 (td, 1H, J_(4′,5′)=4.2, J_(4′,3′)=3.4,H-4′); 4.09 (ddd, 1H, J_(3′,2′)=5.0, J_(3′,OH)=4.8, J_(3′,4′)=3.4,H-3′); 4.41 (ddd, 1H, J_(2′,OH)=6.5, =6.3, J_(2′,3′)=5.0, H-2′); 5.14(d, 1H, J_(OH,3′)=4.8, OH-3′); 5.30 (dd, 1H, J_(OH,5′)=6.5, 4.8, OH-5′);5.34 (d, 1H, J_(OH,2′)=6.5, OH-5′); 6.03 (d, 1H, J_(1′,2′)=6.3, H-1′);7.14 (bs, 1H, NH_(a)H_(b)); 7.50 (d, 1H, J_(5,NH)=2.0, J_(5,2)=0.8,H-5-imidazole); 7.68 (s, 1H, H-6); 7.81 (d, 1H, J_(2,5)=0.8,H-2-imidazole); 8.01 (s, 1H, H-2); 9.99 (bs, 1H, NH_(a)H_(b)); 12.31(bs, 1H, NH-imidazole). ¹³C NMR (125.7 MHz, DMSO-d₆): δ2.11 (CH₂-5′);70.87 (CH-3′); 73.76 (CH-2′); 85.23 (CH-4′); 87.30 (CH-1′); 100.77(C-4a); 110.84 (C-5); 111.81 (CH-5-imidazole); 117.90 (CH-6); 134.95(CH-2-imidazole); 135.10 (C-4-imidazole); 150.56 (C-7a); 152.08 (CH-2);158.59 (C-4). MS (ESI) m/z 333 (M+H), 355 (M+Na). HRMS (ESI) forC₁₄H₁₇N₆O₄ [M+H] calcd: 333.1306. found: 333.1306. Anal. Calcd forC₁₄H₁₆N₆O₄: C, 50.60; H, 4.85; N, 25.29. Found: C, 50.55; H, 5.01; N,24.28.

Example 194-Amino-5-(1H-imidazol-2-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(2s)

To a stirred solution of 1-(N,N-dimethylsulfamoyl)-1H-imidazole (875 mg,5 mmol) in THF (15 ml) was dropwise added n-butyllithium (1.6 M inhexane, 3.1 ml, 5 mmol) at −78° C. during 30 min. Then a solution ofZnCl₂ (1M in THF, 10 ml, 10 mmol) was dropwise added at −78° C. and themixture was warmed to RT during 45 min. Resulting orange solution wastransferred to an argon purged mixture of2′,3′,5′-tri-O-TBS-7-iodotubericidine {compound 4 from Example 17, Step1} (735 mg, 1 mmol) and Pd(PPh₃)₄ (116 mg, 0.1 mmol) and the mixture wasstirred at 90° C. for 24 h. The mixture was diluted with AcOEt (50 ml)and washed with aq EDTA (sat., 25 ml). Aqueous layer was re-extractedwith AcOEt (10 ml). Collected organic extracts were dried over MgSO₄,evaporated and the residue was chromatographed on silica (hexanes/AcOEt,5:1 →1:1; then AcOEt) affording product in a mixture with unseparablestarting protected imidazole. This mixture was directly deprotected bystirring in aq HCl (1M, 2 ml) and MeOH (2 ml) at 100° C. for 24 h.Volatiles were removed in vacuo and the rest was co-evaporated withwater (5×). The residue was treated with MeOH and undissolved solid(imidazole) was filtered off and washed with MeOH. Filtrate wasevaporated and reverse phase chromatography (0-4100% MeOH in water)afforded crude product, which was yet re-purified by columnchromatography on silica (8% MeOH in CHCl₃) providing title compound 2sas yellowish foam (42 mg, 13%). Compound was recrystallized fromMeOH/water. Mp 153-156° C. [α]_(D) −72.1 (c 0.19, DMSO). ¹H NMR (499.8MHz, DMSO-d₆): 3.56 (ddd, 1H, J_(gem)=11.9, J_(5′b,OH)=6.5, =4.6,H-5′b); 3.64 (dd, 1H, J_(gem)=11.9, J_(5′a,OH)=5.0, J_(5′a,4′)=4.1,H-5′a); 3.92 (ddd, 1H, J_(4′,5′)=4.6, 4.1, J_(4′,3′)=3.4, H-4); 4.10(ddd, 1H, J_(3′,OH)=5.2, J_(3′,2′)=5.0, J_(3′,4′)=3.4, H-3′); 4.36 (ddd,1H, J_(2′,OH) 6.3, J_(2′,1′)6.0, J_(2′,3′)=5.0, H-2′); 5.16 (dd, 1H,J_(OH,5′)=6.5, 5.0, OH-5′); 5.17 (d, 1H, J_(OH,3′)=5.2, OH-3′); 5.39 (d,1H, J_(OH,2′)=6.3, OH-5′); 6.05 (d, 1H, J_(1′,2′)=6.0, H-1′); 7.00 (t,1H, J_(4,5)=J_(4,NH)=1.4, H-4-imidazole); 7.22 (dd, 1H, J_(5,NH)=2.0,J_(5,4)=1.4, H-5-imidazole); 7.24 (bs, 1H, NH_(a)H_(b)); 7.86 (s, 1H,H-6); 8.06 (s, 1H, H-2); 10.23 (bs, 1H, NH_(a)H_(b)); 12.42 (bs, 1H,NH-imidazole). ¹³C NMR (125.7 MHz, DMSO-d₆): δ2.25 (CH₂-5′); 70.90(CH-3′); 73.99 (CH-2′); 85.27 (CH-4′); 87.33 (CH-1′); 100.12 (C-4a);107.87 (C-5); 116.81 (CH-5-imidazole); 119.44 (CH-6); 127.22(CH-4-imidazole); 142.49 (C-2-imidazole); 150.82 (C-7a); 152.91 (CH-2);158.58 (C-4). MS (ESI) m/z 333 (M+H), 355 (M+Na). HRMS (ESI) forC₁₄H₁₇N₆O₄ [M+H] calcd: 333.1306. found: 333.1306. Anal. Calcd forC₁₄H₁₆N₆O₄.1.85H₂O.0.55CH₃OH: C, 45.60; 1-1, 5.76; N, 21.93. Found: C,45.67; H, 5.66; N, 21.86.

Example 204-Amino-7-(2-C-methyl-β-D-ribofuranosyl)-5-phenyl-7H-pyrrolo[2,3-d]pyrimidine(7a)

Step 1.4-Chloro-5-iodo-7-(2-C-methyl-2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(10)

To a mixture of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine 8 (903 mg,3.23 mmol), 2-C-methyl-1,2,3,5-tetra-O-benzoyl-β-D-ribofuranose 9 (1.7g, 2.93 mmol) and DBU (1.3 ml, 8.69 mmol) in MeCN (20 ml) was dropwiseadded TMSOTf (2.1 ml, 11.62 mmol) at 0° C. and the mixture was thenstirred at 70° C. for 22.5 h. After cooling, the mixture was dilutedwith AcOEt (100 ml), washed with aq NaHCO₃ (sat., 25 ml), water (25 ml)and brine (25 ml). The organic layer was dried over MgSO₄ andevaporated. The residue was chromatographed on silica (hexanes/toluene,1:1; then hexanes/toluene/MeCN, 49:49:2→3:3:4) affording title compound10 as a white foam (1.04 g, 48%). Compound was recrystallized from EtOH.Mp 95° C. [α]²⁰ _(D) −69.3 (c 0.280, CHCl₃). ¹H NMR (500 MHz, CDCl₃):1.59 (s, 3H, CH₃); 4.72 (td, 1H, J_(4′,3′)=J_(4′,5′b)=5.8,J_(4′,5′a)=3.4, H-4′); 4.85 (dd, 1H, J_(gem)=12.2, J_(5′b,4′)=5.8,H-5′b); 4.95 (dd, 1H, J_(gem)=12.2, J_(5′a,4′)=3.4, H-5′a); 6.03 (d, 1H,J_(3′,4′)=5.8, H-3′); 6.95 (s, 1H, H-1′); 7.34, 7.46 and 7.47 (3×m,3×2H, H-m-Bz); 7.54, 7.59 and 7.61 (3×m, 3×1H, H-p-Bz); 7.69 (s, 1H,H-6); 7.96, 8.10 and 8.11 (3×m, 3×2H, H-o-Bz); 8.75 (s, 1H, H-2). ¹³CNMR (125.7 MHz, CDCl₃): 17.92 (CH₃); 52.68 (C-5); 63.33 (CH₂-5′); 75.55(CH-3′); 80.04 (CH-4′); 84.93 (C-2′); 88.95 (CH-1′); 117.71 (C-4a);128.49, 128.54 and 128.63 (CH-m-Bz); 128.65, 129.50 and 129.61 (C-i-Bz);129.78, 129.83 and 129.92 (CH-o-Bz); 132.66 (CH-6); 133.38, 133.66,133.72 (CH-p-Bz); 150.68 (C-7a); 151.17 (CH-2); 153.15 (C-4); 165.09,165.33 and 166.32 (CO). IR (CHCl₃): 3092, 3066, 3034, 1727, 1602, 1587,1577, 1538, 1504, 1493, 1451, 1444, 1339, 1316, 1269, 1248, 1178, 1162,1141, 1116, 1070, 1027, 1002, 952, 943, 843, 822, 725, 712, 686, 617,600. MS (FAB): m/z 738 (M+H). HRMS (FAB) for C₃₃H₂₆ClIN₃O₇ [M+H] calcd:738.0504. found: 738.0491. Anal. Calcd for C₃₃H₂₅ClIN₃O₇: C, 53.71; H,3.41; N, 5.69. Found: C, 53.91; H, 3.29; N, 5.38.

Step 2.4-Amino-5-iodo-7-(2-C-methyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(6)

A mixture of compound 10 from Step 1 (200 mg, 0.27 mmol) and aq ammonia(25% w/w, 3 ml) in dioxane (3 ml) was stirred in a sealed tube at 120°C. for 10 h. After cooling the volatiles were evaporated and crudeproduct was purified by chromatography on silica (CHCl₃→CHCl₃MeOH, 8:2)and then re-purified by reverse phase chromatography (0→100% MeOH inwater) to afford title compound 6 as white solid (76 mg, 69%). Compoundwas recrystallized from MeOH/MeCN. Mp 207° C. [α]²⁰ _(D) −39.0 (c 0.274,MeOH). ¹H NMR (600 MHz, DMSO-d₆): 0.67 (s, 3H, CH₃); 3.65 (ddd, 1H,J_(gem)=12.1, J_(5′b,OH)=4.8, J_(5′b,4′)=2.7, H-5′b); 3.81 (ddd, 1H,J_(gem)=12.1, J_(5′a,OH)=4.8, J_(5′a,4′)=2.0, H-5′a); 3.79 (ddd, 1H,J_(4′,3′)=9.1, J_(4,5)=2.7, 2.0, H-4′); 3.93 (bd, 1H, J_(3′,4′)=9.1,H-3′); 5.14 (s, 1H, OH-2′); 5.16 (bs, 1H, OH-3′); 5.22 (t, 1H,J_(OH,5′)=4.8, OH-5′); 6.10 (s, 1H, H-1′); 6.67 (bs, 2H, NH₂); 7.82 (s,1H, H-6); 8.10 (s, 1H, H-2). ¹³C NMR (151 MHz, DMSO-d₆): 19.89 (CH₃);51.68 (C-5); 59.46 (CH₂-5′); 71.76 (CH-3′); 78.87 (C-2′); 82.39 (CH-4′);90.71 (CH-1′); 103.11 (C-4a); 126.81 (CH-6); 149.88 (C-7a); 152.23(CH-2); 157.43 (C-4). IR (Mk): 3474, 3429, 3392, 3366, 1631, 1582, 1553,1504, 1440, 1343, 1295, 1147, 1128, 1070, 1045, 789. MS (FAB): m/z 407(M+H). HRMS (FAB) for C₁₂H₁₆H₄O₄ [M+H] calcd: 407.0216. found: 407.0225.Anal. Calcd for C₁₂H₁₅IN₄O₄: C, 35.48; H, 3.72; N, 13.79. Found: C,35.37; H, 3.72; N, 13.39.

Step 3.4-Amino-7-(2-C-methyl-β-D-ribofuranosyl)-5-phenyl-7H-pyrrolo[2,3-d]pyrimidine(7a)

An argon purged mixture of compound 6 from Step 2 (49 mg, 0.12 mmol),phenylboronic acid (25 mg, 0.20 mmol), Na₂CO₃ (144 mg, 1.36 mmol), TPPTS(15.5 mg, 0.027 mmol) and Pd(OAc)₂ (1.4 mg, 6.2 μmol) in water/MeCN(2:1, 1.8 ml) was stirred at 80° C. for 1 h. After cooling, volatileswere removed by evaporation and the residue was purified by reversephase chromatography (0→100% MeOH in water) affording title compound 7aas white solid (30 mg, 70%). Mp 129° C. [α]²⁰ _(D) −55.7 (c 0.226,MeOH). ¹H NMR (600 MHz, DMSO-d₆): 0.75 (s, 3H, CH₃); 3.65 (bdd, 1H,J_(gem)=12.2, J_(5′b,4′)=2.9, H-5′b); 3.82 (bdd, 1H, J_(gem)=12.2,J_(5′a,4′)=2.1, H-5′a); 3.86 (ddd, 1H, J_(4′,3′)=9.1, J_(4′,5′)=2.9,2.1, H-4′); 4.02 (d, 1H, J_(3′,4′)=9.1, H-3′); 5.15 (bs, 3H,OH-2′,3′,5′); 6.10 (bs, 2H, NH₂); 6.23 (s, 1H, H-1′); 7.36 (m, 1H,H-p-Ph); 7.44-7.50 (m, 4H, H-o,m-Ph); 7.70 (s, 1H, H-6); 8.16 (s, 1H,H-2). ¹³C NMR (151 MHz, DMSO-d₆): 20.00 (CH₃); 59.60 (CH₂-5′); 72.01(CH-3′); 78.88 (C-2′); 82.39 (CH-4′); 90.51 (CH-1′); 100.09 (C-4a);116.41 (C-5); 120.75 (CH-6); 127.05 (CH-p-Ph); 128.64 (CH-o-Ph); 129.23(CH-m-Ph); 134.89 (C-i-Ph); 150.67 (C-7a); 151.94 (CH-2); 157.49 (C-4).IR (KBr): 3480, 3431, 3400, 1631, 1622, 1585, 1566, 1537, 1489, 1464,1445, 1299, 1178, 1123, 1073, 1058, 1029, 799, 763, 705, 550. MS (FAB)m/z 357 (M+H). HRMS (FAB) for C₁₈H₂₁N₄O₄: [M+H] calcd: 357.1563. found:357.1557. Anal. Calcd for C₁₈H₂₀N₄O₄.1.6H₂O: C, 56.13; H, 6.07; N,14.54. Found: C, 56.52; H, 5.74; N, 14.14.

Example 214-Amino-5-(4-methoxyphenyl)-7-(2-C-methyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(7b)

Title compound was prepared by following the procedure in Example 20,Step 3. White solid. Yield 67%. Mp 127° C. [α]²⁰ _(D) −48.4 (c 0.225,MeOH). ¹H NMR (600 MHz, DMSO-d₆): 0.74 (s, 3H, CH₃); 3.64 (bdd, 1H,J_(gem)=12.21, J_(5′b,4′)=2.9, H-5′b); 3.80 (s, 3H, CH₃O); 3.81 (bdd,1H, J_(gem)=12.1, J_(5′a,4′)=2.1, H-5′a); 3.85 (ddd, 1H, J_(4′,3′)=9.1,J_(4′,5′)=2.9, 2.1, H-4′); 4.01 (d, 1H, J_(3′,4′)=9.1, H-3); 5.13 (bs,3H, OH-2′,3′,5′); 6.08 (bs, 2H, NH₂); 6.22 (s, 1H, H-1′); 7.04 (m, 2H,H-m-C₆H₄OMe); 7.36 (m, 2H, H-o-C₆H₄OMe); 7.60 (s, 1H, H-6); 8.14 (s, 1H,H-2). ¹³C NMR (151 MHz, DMSO-d₆): 19.99 (CH₃); 55.39 (CH₃O); 59.61(CH₂-5); 72.03 (CH-3); 78.87 (C-2′); 82.34 (CH-4′); 90.47 (CH-1′);100.31 (C-4a); 114.65 (CH-m-C₆H₄OMe); 116.06 (C-5); 120.14 (CH-6);127.04 (C-i-C₆H₄OMe); 129.91 (CH-o-C₆H₄OMe); 150.45 (C-7a); 151.85(CH-2); 157.51 (C-4); 158.58 (C-p-C₆H₄OMe). IR (KBr): 3435, 2836, 1631,1622, 1586, 1565, 1539, 1506, 1464, 1419, 1293, 1247, 1174, 1110, 1072,1033, 839, 798, 791, 712, 550. MS (FAB) m/z 387 (M+H). HRMS (FAB) forC₁₉H₂₃N₄O₅ [M+H] calcd: 387.1668. found: 387.1665. Anal. Calcd forC₁₉H₂₂N₄O₅.1.6H₂O: C, 54.96; H, 6.12; N, 13.49. Found: C, 55.30; H,5.91; N, 13.18.

Example 224-Amino-7-(2-C-methyl-β-D-ribofuranosyl)-5-(naphtalen-1-yl)-7H-pyrrolo[2,3-d]pyrimidine(7c)

Title compound was prepared by following the procedure in Example 20,Step 3. Crude product was prepurified by chromatography on silica (0→20%MeOH in CHCl₃) before final reverse phase chromatography. Tan solid.Yield 41%. Mp 142° C. [α]²⁰ _(D) −58.6 (c 0.239, MeOH). ¹H NMR (500 MHz,DMSO-d₆, T=353 K): 0.90 (s, 3H, CH₃); 3.67 (dd, 1H, J_(gem)=12.2,J_(5′b,4′)=3.5, H-5′b)); 3.83 (dd, 1H, J_(gem)=12.2, J_(5′a,4′)=2.3,H-5′a); 3.91 (ddd, 1H, J_(4′,3′)=8.9, J_(4′,5′)=3.5, 2.3, H-4′); 4.04(d, 1H, J_(3′,4′)=8.9, H-3′); 4.89 (bs, 3H, OH-2′,3′,5′); 5.39 (bs, 2H,NH₂); 6.34 (s, 1H, H-1′); 7.500 (ddd, 1H, J_(7,8)=8.3, J_(7,6)=6.9,J_(7,5)=1.3, H-7-naphth); 7.502 (dd, 1H, J_(2,3)=6.9, J_(2,4)=1.3,H-2-naphth); 7.56 (ddd, 1H, J_(6,5)=8.1, J_(6,7)=6.9, J_(6,8)=1.3,H-6-naphth); 7.60 (dd, 1H, J_(3,4)=8.3, J_(3,2)=6.9, H-3-naphth); 7.63(s, 1H, H-6); 7.75 (dddd, 1H, J_(8,7)=8.3, J_(8,6)=1.3, J_(8,4)=1.0,J_(8,5)=0.8, H-8-naphth); 7.98 (ddd, 1H, J_(4,3)=8.3, J_(4,2)=1.3,J_(4,8)=1.0, H-4-naphth); 8.01 (ddd, 1H, J_(5,6)=8.1, J_(5,7)=1.3,J_(5,8)=0.8, H-5-naphth); 8.19 (s, 1H, H-2). ¹³C NMR (151 MHz, DMSO-d₆,T=353 K): 19.73 (CH₃); 59.70 (CH₂-5′); 72.30 (CH-3′); 78.69 (C-2′);82.28 (CH-4′); 90.64 (CH-1′); 102.10 (C-4a); 113.02 (C-5); 121.50(CH-6); 125.27 (CH-8-naphth); 125.38 (CH-3-naphth); 125.98(CH-6-naphth); 126.42 (CH-7-naphth); 127.82 (CH-4-naphth); 128.12(CH-2,5-naphth); 131.63 (C-1-naphth); 132.13 (C-8a-naphth); 133.39(C-4a-naphth); 150.05 (C-7a); 151.61 (CH-2); 156.96 (C-4). IR (KBr):3478, 3438, 3395, 3058, 2973, 1620, 1583, 1578, 1568, 1533, 1507, 1468,1398, 1377, 1298, 1256, 1178, 1143, 1117, 1071, 1050, 1035, 1018, 852,799, 786, 780, 740. MS (ESI) m/z 407 (M+H), 429 (M+Na). HRMS (ESI) forC₂₂H₂₃N₄O₄ [M+H] calcd: 407.1714. found: 407.1704. Anal. Calcd forC₂₂H₂₂N₄O₄.1.5H₂O: C, 60.96; H, 5.81; N, 12.93. Found: C, 61.30; H,5.72; N, 13.28.

Example 234-Amino-7-(2-C-methyl-β-D-ribofuranosyl)-5-(naphtalen-2-yl)-7H-pyrrolo[2,3-d]pyrimidine(7d)

Title compound was prepared by following the procedure in Example 20,Step 3. Crude product was prepurified by chromatography on silica (0→20%MeOH in CHCl₃) before final reverse phase chromatography. Cream solid.Yield 65%. Mp 143° C. [α]²⁰ _(D) −64.3 (c 0.253, MeOH). ¹H NMR (500 MHz,DMSO-d₆): 0.79 (s, 3H, CH₃); 3.66 (ddd, 1H, J_(gem)=12.6, J_(5′b,OH)⁼5.0, J_(5′b,4′)=2.9, H-5′b); 3.84 (ddd, 1H, J_(gem)=12.6,J_(5′a,OH)=5.0, J_(5′a,4′)=2.1, H-5′a); 3.88 (ddd, 1H, J_(4′,3′)=9.1,J_(4′,5′)=2.9, 2.1, H-4′); 4.06 (bdd, 1H, J_(3′,4′)=9.1, J_(3′,OH)=4.6,H-3′); 5.11-5.17 (bm, 3H, OH-2′,3′,5′); 6.19 (bs, 2H, NH₂); 6.27 (s, 1H,H-1′); 7.52 (m, 1H, H-6-naphth); 7.55 (m, 1H, H-7-naphth); 7.62 (dd, 1H,J_(3,4)=8.5, J_(3,1)=1.8, H-3-naphth); 7.81 (s, 1H, H-6); 7.94-7.97 (m,3H, H-1,5,8-naphth); 8.01 (d, 1H, J_(4,3)=8.5, H-4-naphth); 8.18 (s, 1H,H-2). ¹³C NMR (125.7 MHz, DMSO-d₆): 20.03 (CH₃); 59.66 (CH₂-5′); 72.08(CH-3′); 78.90 (C-2′); 82.41 (CH-4′); 90.57 (CH-1′); 100.24 (C-4a);116.45 (C-5); 121.14 (CH-6); 126.10 (CH-6-naphth); 126.76 (CH-7-naphth);126.82 (CH-1-naphth); 127.23 (CH-3-naphth); 127.87 and 128.02(CH-5,8-naphth); 128.70 (CH-4-naphth); 132.00 (C-4a-naphth); 132.35(C-1-naphth); 133.46 (C-8a-naphth); 150.82 (C-7a); 152.02 (CH-2); 157.61(C-4). IR (KBr): 3506, 3481, 3452, 3402, 3325, 3242, 3210, 3110, 3052,2973, 1645, 1624, 1605, 1587, 1567, 1535, 1503, 1469, 1378, 1298, 1274,1142, 1128, 1116, 1075, 1057, 1050, 1019, 897, 860, 821, 796, 767, 750,478. MS (ESI) m/z 407 (M+H), 429 (M+Na). HRMS (ESI) for C₂₂H₂₃N₄O₄ [M+H]calcd: 407.1714. found: 407.1715. Anal. Calcd for C₂₂H₂₂N₄O₄.1.3H₂O: C,61.47; H, 5.77; N, 13.03. Found: C, 61.83; H, 5.51; N, 12.65.

Example 244-Amino-5-(furan-2-yl)-7-(2-C-methyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(7e)

Title compound was prepared by following the procedure in Example 20,Step 3. White solid. Yield 55%. Mp 130° C. [α]²⁰ _(D) −53.3 (c 0.250,MeOH). ¹H NMR (600 MHz, DMSO-d₆): 0.72 (s, 3H, CH₃); 3.68 and 3.85(2×bd, 2H, J_(gem)=12.0, H-5′); 3.87 (ddd, 1H, =9.1, J_(4′,5′)=2.9, 2.0,H-4′); 4.00 (bd, 1H, J_(3′,4′)=9.1, H-3′); 5.16 (bs, 2H, OH-2′,3′); 5.25(bs, 1H, OH-5′); 6.19 (s, 1H, H-1′); 6.58 (dd, 1H, J_(3,4)=3.3,J_(3,5)=0.8, H-3-furyl); 6.60 (dd, 1H, J_(4,3)=3.3, J_(4,5)=1.9,H-4-furyl); 6.90 (bs, 2H, NH₂); 7.78 (dd, 1H, J_(5,4)=1.9, J_(5,3)=0.8,H-5-furyl); 8.01 (s, 1H, H-6); 8.14 (s, 1H, H-2). ¹³C NMR (151 MHz,DMSO-d₆): 19.91 (CH₃); 59.66 (CH₂-5′); 71.92 (CH-3′); 78.85 (C-2′);82.48 (CH-4′); 90.60 (CH-1′); 99.03 (C-4a); 105.22 (CH-3-furyl); 106.10(C-5); 112.13 (CH-4-furyl); 120.07 (CH-6); 142.17 (CH-5-furyl); 148.98(C-2-furyl); 150.60 (C-7a); 152.35 (CH-2); 157.46 (C-4). IR (KBr): 3474,3385, 3351, 3245, 3200, 1630, 1575, 1560, 1531, 1497, 1472, 1455, 1379,1299, 1124, 1072, 1049, 1016, 893, 795, 594. MS (ESI) m/z 347 (M+H), 369(M+Na). HRMS (ESI) for C₁₆H₁₉N₄O₅ [M+H] calcd: 347.1355. found:347.1347. Anal. Calcd for C₁₆H₁₈N₄O₅.0.7H₂O: C, 53.54; H, 5.45; N,15.61. Found: C, 53.86; H, 5.48; N, 15.22.

Example 254-Amino-7-(2-C-methyl-β-D-ribofuranosyl)-5-(thiophen-2-yl)-7H-pyrrolo[2,3-d]pyrimidine(7f)

Title compound was prepared by following the procedure in Example 20,Step 3. White solid. Yield 77%. Mp 131° C. [α]²⁰ _(D) −51.4 (c 0.255,MeOH). ¹H NMR (600 MHz, DMSO-d₆): 0.73 (s, 3H, CH₃); 3.65 (dd, 1H,J_(gem)=12.3, J_(5′b,4′)=2.9, H-5′b); 3.83 (dd, 1H, J_(gem)=12.3,J_(5′a,4′)=2.1, H-5′a); 3.86 (ddd, 1H, J_(4′,3′)=9.1, J_(4′,5′)=2.9,2.1, H-4′); 4.00 (bd, 1H, J_(3′,4′)=9.1, H-3′); 5.20 (bs, 3H,OH-2′,3′,5′); 6.19 (s, 1H, H-1′); 6.30 (bs, 2H, NH₂); 7.13 (dd, 1H,J_(3,4)=3.5, J_(3,5)=1.2, H-3-thienyl); 7.16 (dd, 1H, J_(4,5)=5.2,J_(4,3)=3.5, H-4-thienyl); 7.55 (dd, 1H, J_(5,4)=5.2, J_(5,3)=1.1,H-2-thienyl); 7.82 (s, 1H, H-6); 8.16 (s, 1H, H-2). ¹³C NMR (151 MHz,DMSO-d₆): 19.96 (CH₃); 59.37 (CH₂-5′); 71.76 (CH-3′); 78.88 (C-2′);82.38 (CH-4′); 90.53 (CH-1′); 100.27 (C-4a); 108.51 (C-5); 121.64(CH-6); 125.93 (CH-5-thienyl); 126.42 (CH-3-thienyl); 128.53(CH-4-thienyl); 136.04 (C-2-thienyl); 150.40 (C-7a); 152.26 (CH-2);157.47 (C-4). IR (CH-1′): 3471, 3380, 3350, 3235, 3195, 1622, 1591,1575, 1549, 1507, 1460, 1430, 1372, 1349, 1296, 1228, 1122, 1071, 1050,850, 796, 707, 559. MS (ESI) m/z 363 (M+H), 385 (M+Na). HRMS (ESI) forC₁₆H₁₉N₄O₄S [M+H] calcd: 363.1127. found: 363.1121. Anal. Calcd forC₁₆H₁₈N₄O₄S.0.95H₂O: C, 50.64; H, 5.28; N, 14.76. Found: C, 51.03; H,5.06; N, 14.40.

Example 264-Amino-5-(furan-3-yl)-7-(2-C-methyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(7g)

Title compound was prepared by following the procedure in Example 20,Step 3. Reaction time 2.5 h. White solid. Yield 59%. Mp 123° C. [α]²⁰_(D) −54.5 (c 0.217, MeOH). ¹H NMR (600 MHz, DMSO-d₆): 0.72 (s, 3H,CH₃); 3.65 (dd, 1H, J_(gem)=12.2, J_(5′b,4′)=3.0, H-5′b); 3.82 (dd, 1H,J_(gem)=12.2, J_(5′a,4′)=2.1, H-5′a); 3.85 (ddd, 1H, J_(4′,3′)=9.1,J_(4′,5′)=3.0, 2.1, H-4′); 3.99 (d, 1H, J_(3′,4′)=9.1, H-3′); 5.16 (bs,3H, OH-2′,3′,5′); 6.19 (s, 1H, H-1′); 6.25 (bs, 2H, NH₂); 6.66 (dd, 1H,J_(4,5)=1.7, J_(4′,2′)=0.8, H-4-furyl); 7.65 (s, 1H, H-6); 7.79 (t, 1H,J_(5,2)=J_(5,4)=1.7, H-5-furyl); 7.80 (dd, 1H, J_(2,5)=1.7, J_(2,4)=0.8,H-2-furyl); 8.13 (s, 1H, H-2). ¹³C NMR (151 MHz, DMSO-d₆): 19.99 (CH₃);59.74 (CH₂-5′); 72.08 (CH-3); 78.87 (C-2′); 82.41 (CH-4); 90.56 (CH-1′);100.67 (C-4a); 106.27 (C-5); 111.78 (CH-4-furyl); 119.00 (C-3-furyl);120.61 (CH-6); 139.79 (CH-2-furyl); 144.43 (CH-5-furyl); 150.52 (C-7a);152.03 (CH-2); 157.68 (C-4). IR (KBr): 3478, 3337, 3243, 3208, 3150,1623, 1583, 1561, 1534, 1498, 1455, 1378, 1357, 1350, 1300, 1159, 1120,1071, 1056, 1026, 874, 793, 602. MS (ESI) m/z 347 (M+H). HRMS (ESI) forC₁₆H₁₉N₄O₅ [M+H] calcd: 347.1350. found: 347.1349. Anal. Calcd forC₁₆H₁₈N₄O₅.1.7H₂O: C, 50.98; 1-1, 5.72; N, 14.86. Found: C, 51.30; H,5.33; N, 14.46.

Example 274-Amino-7-(2-C-methyl-β-D-ribofuranosyl)-5-(thiophen-3-yl)-7H-pyrrolo[2,3-d]pyrimidine(7h)

Title compound was prepared by following the procedure in Example 20,Step 3. White solid. Yield 62%. Mp 128° C. [α]²⁰ _(D) −60.3 (c 0.222,MeOH). ¹H NMR (600 MHz, DMSO-d₆): 0.74 (s, 3H, CH₃); 3.65 (ddd, 1H,J_(gem)=12.1, J_(5′b,OH)=4.8, J_(5′b,4′)=2.9, H-5′b); 3.82 (ddd, 1H,J_(gem)=12.1, J_(5′a,OH)=4.8, J_(5′a,4′)=2.1, H-5′a); 3.85 (ddd, 1H,J_(4′,3′)=9.0, J_(4′,5)=2.9, 2.1, H-4′); 4.00 (dd, 1H, J_(3,4)=9.0,J_(3′,OH)=7.1, H-3′); 5.12 (d, 1H, J_(OH,3′)=7.1, OH-3′); 5.13 (s, 1H,OH-2′); 5.16 (t, 1H, J_(OH,5′)=4.8, OH-5′); 6.20 (s, 1H, H-1′); 6.20(bs, 2H, NH₂); 7.24 (dd, 1H, J_(4,5)=4.9, J_(4,2)=1.4, H-4-thienyl);7.49 (dd, 1H, J_(2,5)=2.9, J_(2,4)=1.4, H-2-thienyl); 7.698 (dd, 1H,J_(5,4)=4.9, J_(5,2)=2.9, H-5-thienyl); 7.70 (s, 1H, H-6); 8.14 (s, 1H,H-2). ¹³C NMR (151 MHz, DMSO-d₆): 20.01 (CH₃); 59.67 (CH₂-5′); 72.04(CH-3′); 78.89 (C-2′); 82.41 (CH-4′); 90.55 (CH-1′); 100.44 (C-4a);111.08 (C-5); 120.73 (CH-6); 122.11 (CH-2-thienyl); 127.66(CH-5-thienyl); 128.72 (CH-4-thienyl); 135.16 (C-3-thienyl); 150.38(C-7a); 151.96 (CH-2); 157.57 (C-4). IR (KBr): 3475, 3351, 3240, 3200,3120, 3107, 1621, 1594, 1575, 1549, 1510, 1465, 1409, 1378, 1346, 1297,1139, 1123, 1072, 1045, 861, 788. MS (ESI) m/z 363 (M+H), 385 (M+Na).HRMS (ESI) for C₁₆H₁₉N₄O₄S [M+H] calcd: 363.1122. found: 363.1122. Anal.Calcd for C₁₆H₁₈N₄O₄S.1.1H₂O: C, 50.28; H, 5.33; N, 14.66. Found: C,50.42; H, 5.20; N, 14.45.

Example 284-Amino-5-(benzofuran-2-yl)-7-(2-C-methyl-β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(7i)

Title compound was prepared by following the procedure in Example 20,Step 3. White solid. Yield 66%. Mp 248° C. (dec.). [α]²⁰ _(D) −55.0 (c0.220, MeOH). ¹H NMR (500 MHz, DMSO-d₆): 0.76 (s, 3H, CH₃); 3.72 (ddd,1H, J_(gem)=12.5, J_(5′b,OH)=5.0, J_(5′b,4′)=2.9, H-5′b); 3.88 (ddd, 1H,J_(gem)=12.5, J_(5′a,OH)=5.0, J_(5′a,4′)=2.0, H-5′a); 3.90 (ddd, 1H,J_(4′,3′)=9.2, J_(4′,5′)=2.9, 2.0, H-4′); 4.04 (dd, 1H, J_(3′,4′)=9.2,J_(3′,OH)=7.0, H-3′); 5.17 (d, 1H, J_(OH,3′)=7.0, OH-3′); 5.19 (s, 1H,OH-2′); 5.28 (t, 1H, J_(OH,5′)=5.0, OH-5′); 6.23 (s, 1H, H-1′); 6.98(bs, 2H, NH₂); 7.04 (d, 1H, J_(3,7)=1.0, H-3-benzofuryl); 7.27 (td, 1H,J_(5′,4′)=J_(5,6)=7.3, J_(5,7)=1.4, H-5-benzofuryl); 7.29 (td, 1H,J_(6,5)=J_(6,7)=7.3, J_(6,4)=1.7, H-6-benzofuryl); 7.61-7.67 (m, 2H,H-4,7-benzofuryl); 8.19 (s, 1H, H-2); 8.26 (s, 1H, H-6). ¹³C NMR (125.7MHz, DMSO-d₆): 19.96 (CH₃); 59.72 (CH₂-5′); 71.97 (CH-3′); 78.91 (C-2′);82.59 (CH-4′); 90.74 (CH-1′); 99.19 (C-4a); 101.58 (CH-3-benzofuryl);105.47 (C-5); 111.30 (CH-7-benzofuryl); 120.81 (CH-4-benzofuryl); 122.32(CH-6); 123.73 (CH-5-benzofuryl); 124.08 (CH-6-benzofuryl); 129.01(C-3a-benzofuryl); 150.94 (C-7a); 151.45 (C-2-benzofuryl); 152.58(CH-2); 154.00 (C-7a-benzofuryl); 157.50 (C-4). MS (ESI, negative mode)m/z 395 (M−H). HRMS (ESI, negative mode) for C₂₀H₁₉N₄O₅: [M−H] calcd:395.1350. found: 395.1358. Anal. Calcd for C₂₀H₂₀N₄O₅.⅓H₂O: C, 59.70; H,5.18; N, 13.92. Found: C, 59.98; H, 5.13; N, 13.46.

Example 294-Amino-5-phenyl-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-triphosphate sodium salt (13a)

Step 1. 4-Amino-5-iodo-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-triphosphate sodium salt (11)

Phosphorus oxychloride (45 μl, 0.49 mmol) was dropwise added to astirred mixture of 7-iodotubericidine 1 (150 mg, 0.38 mmol) in trimethylphosphate (1 ml) at 0° C. and the solution was stirred at 0° C. for 1.25h. A freshly prepared solution of bis(tri-n-butylammonium) pyrophosphate(1.05 g, 1.91 mmol) and tri-n-butylamine (0.4 ml, 1.66 mmol) in dry DMF(4 ml) was stirred with molecular sieves at 0° C. for at least 15 minand then added to the stirred reaction mixture at 0° C. The mixture washeld at the same temperature for 1.5 h before being quenched with aqTEAB (2M, 1.2 ml). Volatiles were removed in vacuo and the rest wasseveral times co-evaporated with water. The residue was purified byion-exchange chromatography on DEAE-Sephadex (0→60% 2M aq TEAB in H₂O)and obtained triethylammonium salt of product was converted to sodiumsalt by passing through column of Dowex 50 (Na⁺ form). Lyophilizationafforded title compound 11 as a white cotton (131 mg, 48%). ¹H NMR (500MHz, D₂O+phosphate buffer, pH=7.1, ref_(dioxane)=3.75 ppm): 4.15 (ddd,1H, J_(gem)=11.6, J_(H,P)=4.9, J_(5′b,4′)=3.3, H-5′b); 4.24 (ddd, 1H,J_(gem)=11.6, J_(H,P)=6.6, J_(5′a,4′)=3.2, H-5′a); 4.34 (m, 1H,J_(4′,5′)=3.3, 3.2, J_(4′,3′)=2.9, J_(H,P)=2.1, H-4′); 4.54 (ddd, 1H,J_(3′,2′)=5.3, J_(3′,4′)=2.9, J_(H,P)=0.4, H-3′); 4.65 (dd, 1H,J_(2′,1′)=6.7, J_(2′,3′)=5.3, H-2′); 6.21 (d, 1H, J_(1′,2′)=6.7, H-1′);7.71 (s, 1H, H-8); 8.09 (s, 1H, H-2). ¹³C NMR (125.7 MHz, D₂O+phosphatebuffer, pH=7.1, ref_(dioxane)=69.3 ppm): 55.01 (C-7); 68.18 (d,J_(C,P)=6, CH₂-5′); 73.21 (CH-3′); 76.51 (CH-2′); 86.33 (d, J_(C,P)=9,CH-4′); 88.50 (CH-1′); 106.76 (C-4a); 129.78 (CH-6); 152.52 (C-7a);154.17 (CH-2); 159.72 (C-4). ³¹P (¹H dec.) NMR (202.4 MHz, D₂O+phosphatebuffer, pH=7.1, ref_(H3PO4)=0 ppm): −20.78 (t, J=19.5, P_(β)); −9.66 (d,J=19.5, P_(α)); −7.00 (d, J=19.5, P_(γ)). MS (ESI) m/z 699 (M+H), 721(M+Na). MS (ESI, negative mode) m/z 631 (M−3Na+2H), 653 (M−2Na+H), 675(M−Na). HRMS (ESI, negative mode)⁻ for C₁₁H₁₄IN₄NaO₁₃P₃ [M−2Na+H] calcd:652.8713. found: 652.8731.

Step 2.4-Amino-5-phenyl-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-triphosphate sodium salt (13a)

An an argon purged mixture of Pd(OAc)₂ (1.5 mg, 6.7 μmol) and TPPTS(17.3 mg, 30 μmol) in water/MeCN (2:1, 1.2 ml) was sonicated to fulldissolution and quarter of this pre-prepared solution (0.3 ml, ¼ oftotal amount) was added to an argon purged mixture of compound 11 fromStep 1 (20.1 mg, 29 μmol), phenylboronic acid (5.2 mg, 42 μmol), Cs₂CO₃(27 mg, 83 μmol) in water/MeCN (2:1, 0.6 ml) and the mixture was stirredat 110° C. for 30 min. After cooling, the mixture was filtered throughmicrofilter and separated by HPLC on C-18 phase (0→100% MeOH in 0.1M aqTEAB) affording after ion exchange on Dowex 50 (Na⁺ form) andlyophilization title compound 13a as white cotton (8.6 mg, 46%). ¹H NMR(500 MHz, D₂O+phosphate buffer, pH=7.1, ref_(dioxane)=3.75 ppm): 4.14(ddd, 1H, J_(gem)=11.6, J_(H,P)=4.7, J_(5′b,4′)=3.5, H-5′b); 4.25 (ddd,1H, J_(gem)=11.6, J_(H,P)=6.5, =3.3, H-5′a); 4.35 (m, 1H, J_(4′,5′)=3.5,3.3, =2.9, J_(H,P)=1.7, H-4′); 4.57 (dd, 1H, J_(3′,2′)=5.4, =2.9, H-3′);4.73 (dd, 1H, J_(2′,1′)=6.9, J_(H,P)=5.4, H-2′); 6.30 (d, 1H,J_(1′,2′)=6.9, H-1′); 7.45 (m, 1H, H-p-Ph); 7.49-7.56 (m, 4H, H-o,m-Ph);7.57 (s, 1H, H-6); 8.16 (s, 1H, H-2). ¹³C NMR (125.7 MHz, D₂O+phosphatebuffer, pH=7.1, ref_(dioxane)=69.3 ppm): 68.22 (d, J_(C,P)=5, CH₂-5′);73.26 (CH-3′); 76.24 (CH-2′); 86.31 (d, J_(C,P)=9, CH-4′); 88.34(CH-1′); 103.75 (C-4a); 121.56 (C-5); 122.86 (CH-6); 130.41 (CH-p-Ph);131.59 and 131.94 (CH-o,m-Ph); 136.10 (C-i-Ph); 153.15 (C-7a); 153.67(CH-2); 159.61 (C-4). ³¹P (¹H dec.) NMR (202.4 MHz, D₂O+phosphatebuffer, pH=7.1, ref_(H3PO4)=0 ppm): −21.32 (dd, J=19.3, 19.0, P_(β));−10.45 (d, J=19.3, P_(α)); −6.98 (d, J=19.0, P_(γ)). MS (ESI, negativemode) m/z 581 (M−3Na+2H), 603 (M−2Na+H), 625 (M−Na). HRMS (ESI, negativemode) for C₁₇H₂₀N₄O₁₃P₃ [M−3Na+2H] calcd: 581.0240. found: 581.0253.

Example 304-Amino-5-(4-fluorophenyl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-triphosphate sodium salt (13b)

Title compound was prepared by following the procedure in Example 29,Step 2. White cotton. Yield 25%. ¹H NMR (500 MHz, D₂O+phosphate buffer,pH=7.1, ref_(dioxane)=3.75 ppm): 4.13 (dt, 1H, J_(gem)=11.4,J_(H,P)p=J_(5′b,4′)=4.2, H-5′b); 4.25 (ddd, 1H, J_(gem)=11.4,J_(H,P)=6.5, =3.3, H-5′a); 4.35 (bddd, 1H, J_(4′,5′)=4.2, 3.3,J_(4′,3′)=2.7, H-4′); 4.58 (dd, 1H, =5.4, J_(3′,4′)=2.7, H-3′); 4.72(dd, 1H, J_(2′,1′)=6.9, =5.4, H-2); 6.31 (d, 1H, J_(1′,2′)=6.9, H-1′);7.24 (m, 2H, H-m-C₆H₄F); 7.53 (m, 2H, H-o-C₆H₄F); 7.56 (s, 1H, H-6);8.17 (s, 1H, H-2). ¹³C NMR (125.7 MHz, D₂O+phosphate buffer, pH=7.1,ref_(diaxane)=69.3 ppm): 68.19 (d, J_(C,P)=6, CH₂-5′); 73.26 (CH-3′);76.21 (CH-2′); 86.34 (d, J_(C,P)=9, CH-4); 88.26 (CH-1′); 103.93 (C-4a);118.57 (d, J_(C,P)=22, CH-m-C₆H₄F); 120.54 (C-5); 122.83 (CH-6); 132.26(d, J_(C,P)=3, C-i-C₆H₄F); 133.44 (d, J_(C,P)=8, CH-o-C₆H₄F); 153.24(C-7a); 154.16 (CH-2); 159.98 (C-4); 164.95 (d, J_(C,F)=244, C-p-C₆H₄F).³¹P (¹H dec.) NMR (202.4 MHz, D₂O+phosphate buffer, pH=7.1,ref_(H3PO4)=0 ppm): −21.04 (dd, J=18.9, 18.4, P_(β)); −10.39 (d, J=18.9,P_(α)); −6.12 (d, J=18.4, P_(γ)). MS (ESI, negative mode) m/z 621(M−2Na+H), 643 (M−Na). HRMS (ESI, negative mode) for C₁₇H₁₇FN₄Na₂O₁₃P₃[M−Na] calcd: 642.9779. found: 642.9789.

Example 314-Amino-5-(furan-2-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-triphosphate sodium salt (13c)

Title compound was prepared by following the procedure in Example 29,Step 2. Re-purification by ion-exchange HPLC on Poros HQ (TEAB gradient)after HPLC on C-18. White cotton. Yield 27%. ¹H NMR (500 MHz,D₂O+phosphate buffer, pH=7.1, ref_(dioxane)=3.75 ppm): 4.19 (ddd, 1H,J_(gem)=11.7, J_(H,P)=4.7, J_(5′b,4′)=3.2, H-5′b); 4.28 (ddd, 1H,J_(gem)=11.7, J_(H,P)=6.2, J_(5′a,4′)=2.8, H-5′a); 4.36 (ddd, 1H,J_(4′,5′)=3.2, 2.8, J_(4′,3′)=2.9, H-4′); 4.59 (dd, 1H, J_(3,2)=5.2,=2.9, H-3′); 4.71 (dd, 1H, =6.6, J_(2′,3′)=5.2, H-2′); 6.25 (d, 1H,J_(1′,2′)=6.6, H-1′); 6.57 (dd, 1H, J_(4,3)=3.4, J_(4,5)=1.9,H-4-furyl); 6.75 (dd, 1H, J_(3,4)=3.4, J_(3,5)=0.7, H-3-furyl); 7.57(dd, 1H, J_(5,4)=1.9, J_(5,3)=0.7, H-5-furyl); 7.84 (s, 1H, H-6); 8.08(s, 1H, H-2). ¹³C NMR (125.7 MHz, D₂O+phosphate buffer, pH=7.1,ref_(dioxane)=69.3 ppm): 68.14 (d, J_(C,P)=5, CH₂-5′); 73.28 (CH-3′);76.60 (CH-2′); 86.40 (d, J_(C,P)=9, CH-4′); 88.60 (CH-1′); 102.45(C-4a); 108.95 (CH-3-furyl); 111.18 (C-5); 114.80 (CH-4-furyl); 122.05(CH-6); 144.63 (CH-5-furyl); 150.34 (C-2-furyl); 152.81 (C-7a); 153.46(CH-2); 159.28 (C-4). ³¹P (¹H dec.) NMR (202.4 MHz, D₂O+phosphatebuffer, pH=7.1, ref_(H3PO4)=0 ppm): −21.09 (dd, J=19.2, 18.7, P_(β));−10.43 (d, J=19.2, P_(α)); −6.25 (d, J=18.7, P_(γ)). MS (ESI, negativemode) m/z 593 (M−2Na+H), 615 (M−Na). HRMS (ESI, negative mode) forC₁₅H₁₇N₄NaO₁₄P₃ [M−2Na+H] calcd: 592.9846. found: 592.9868; forC₁₅H₁₆N₄Na₂O₁₄P₃ [M−Na] calcd: 614.9666. found: 614.9686; forC₁₅H₁₄N₄Na₄O₁₄P₃ [M−2H+Na] calcd: 658.9310. found: 658.9321.

Example 324-Amino-7-(β-D-ribofuranosyl)-5-(thiophen-2-yl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-triphosphate sodium salt (13d)

Title compound was prepared by following the procedure in Example 29,Step 2. White solid. Yield 53%. ¹H NMR (500 MHz, D₂O+phosphate buffer,pH=7.1, ref_(dioxane)=3.75 ppm): 4.14 (ddd, 1H, J_(gem)=11.7,J_(H,P)=4.9, J_(5′b,4′)=3.6, H-5′b); 4.28 (ddd, 1H, J_(gem)=11.7,J_(H,P)=6.9, J_(5′a,4′)=3.6, H-5′a); 4.35 (td, 1H, J_(4′,5′)=3.6,J_(4′,3′)=2.7, H-4′); 4.58 (dd, 1H, J_(3′,2′)=5.3, J_(3′,4′)=2.7, H-3′);4.73 (dd, 1H, J_(2′,1′)7.1, J_(2′,3′)=5.3, H-2′); 6.30 (d, 1H,J_(1′,2′)=7.1, H-1′); 7.21 (dd, 1H, J_(4,5)=5.0, J_(4,3)=3.5,H-4-thienyl); 7.22 (dd, 1H, J_(3,4)=3.5, J_(3,5)=1.4, H-3-thienyl); 7.51(dd, 1H, J_(5,4)=5.0, J_(5,3)=1.4, H-5-thienyl); 7.64 (s, 1H, H-6); 8.18(s, 1H, H-2). ¹³C NMR (125.7 MHz, D₂O+phosphate buffer, pH=7.1,ref_(dioxane)=69.3 ppm): 68.20 (d, J_(C,P)=5, CH₂-5′); 73.22 (CH-3′);76.21 (CH-2′); 86.40 (d, J_(C,P)=9, CH-4); 88.28 (CH-1′); 104.13 (C-4a);113.58 (C-5); 123.93 (CH-6); 129.26 (CH-5-thienyl); 130.14(CH-3-thienyl); 130.95 (CH-4-thienyl); 137.11 (C-2-thienyl); 153.14(C-7a); 154.52 (CH-2); 160.04 (C-4). ³¹P (¹H dec.) NMR (202.4 MHz,D₂O+phosphate buffer, pH=7.1, ref_(H3PO4)=0 ppm): −21.12 (bdd, J=20.5,19.4, P_(β)); −10.41 (d, J=19.4, P_(α)); −6.23 (d, J=20.5, P_(γ)). MS(ESI, negative mode) m/z 609 (M−2Na+H), 631 (M−Na). HRMS (ESL negativemode) for C₁₅H₁₄N₄NaO₁₃P₃S [M−2Na+H] calcd: 608.9618. found: 608.9637.

Example 334-Amino-5-(furan-3-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-triphosphate sodium salt (13e)

Title compound was prepared by following the procedure in Example 29,Step 2. Re-purification by ion-exchange HPLC on Poros HQ (TEAB gradient)after HPLC on C-18. Pale yellow solid. Yield 37%. ¹H NMR (600 MHz,D₂O+phosphate buffer, pH=7.1, ref_(dioxane)=3.75 ppm): 4.14 (ddd, 1H,J_(gem)=11.2, J_(H,P)=4.9, J_(5′b,4′)=2.9, H-5′b); 4.25 (ddd, 1H,J_(gem)=11.2, J_(H,P)=6.4,J_(5′a,4′)=3.1, H-5′a); 4.35 (ddd, 1H,J_(4′,5′)=3.2, 2.9, J_(4′,3′)=2.4, H-4′); 4.57 (dd, 1H,J_(3′,2′)=5.4,J_(3′,4′)=2.4, H-3′); 4.73 (dd, 1H, J_(2′,1′)=6.9,J_(2′,3′)=5.4, H-2′); 6.29 (d, 1H, J_(1′,2′)=6.9, H-1′); 6.73 (dd, 1H,J_(4,5)=1.9,J_(4,2)=0.9, H-4-furyl); 7.58 (s, 1H, H-6); 7.65 (dd, 1H,J_(5,4)=1.9, J_(5,2)=1.6, H-5-furyl); 7.75 (dd, 1H, J_(2,5)=1.6,J_(2,4)=0.9, H-2-furyl); 8.16 (s, 1H, H-2). ¹³C NMR (151 MHz,D₂O+phosphate buffer, pH=7.1, ref_(dioxane)=69.3 ppm): 68.22 (d,J_(C,P)=5, CH₂-5′); 73.32 (CH-3′); 76.22 (CH-2); 86.35 (d, J_(C,P)=9,CH-4′); 88.24 (CH-1′); 104.39 (C-4a); 111.40 (C-5); 114.18 (CH-4-furyl);120.38 (C-3-furyl); 122.84 (CH-6); 143.14 (CH-2-furyl); 147.03(CH-5-furyl); 153.22 (C-7a); 154.35 (CH-2); 160.22 (C-4). ³¹P (¹H dec.)NMR (202.4 MHz, D₂O+phosphate buffer, pH=7.1, ref_(H3PO4)=0 ppm): −20.54(bdd, J=17.8, 15.0, P_(β)); −10.35 (d, J=17.8, P_(α)); −5.66 (d, J=15.0,P_(γ)). MS (ESI, negative mode) m/z 593 (M−2Na+H), 615 (M−Na). HRMS(ESI, negative mode) for C₁₅H₁₆N₄Na₂O₁₄P₃ [M−Na] calcd: 614.9666. found:614.9678.

Example 344-Amino-7-(β-D-ribofuranosyl)-5-(thiophen-3-yl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-triphosphate sodium salt (13f)

Title compound was prepared by following the procedure in Example 29,Step 2. White cotton. Yield 67%. ¹H NMR (500 MHz, D₂O+phosphate buffer,pH=7.1, ref_(dioxane)=3.75 ppm): 4.14 (ddd, 1H, J_(gem)=11.6,J_(H,P)=4.8, J_(5′b,4′)=3.5, H-5′b); 4.25 (ddd, 1H, J_(gem)=11.6,J_(H,P)=6.6, J_(5′a,4′)=3.2, H-5′a); 4.35 (ddd, 1H, J_(4′,5′)=3.5,3.2,J_(4′,3′)=2.7, H-4′); 4.58 (dd, 1H, J_(3′,2′)=5.3, J_(3′,4′)=2.7,H-3′); 4.74 (dd, 1H, J_(2′,1′)=7.0, J_(2′,3′)=5.3, H-2′); 6.30 (d, 1H,J_(1′,2′)=7.0, H-1′); 7.33 (dd, 1H, J_(4,5)=5.0, J_(4,2)=1.4,H-4-thienyl); 7.52 (dd, 1H, J_(2,5)=3.0, J_(2,4)=1.4, H-2-thienyl);7.607 (s, 1H, H-6); 7.610 (dd, 1H, J_(5,4)=5.0, J_(5,2)=3.0,H-5-thienyl); 8.17 (s, 1H, H-2). ¹³C NMR (125.7 MHz, D₂O+phosphatebuffer, pH=7.1, ref_(dioxane)=69.3 ppm): 68.20 (d, J_(C,P)=5, CH₂-5′);73.30 (CH-3′); 76.24 (CH-2′); 86.38 (d, J_(C,P) 9, CH-4′); 88.28(CH-1′); 104.09 (C-4a); 116.08 (C-5); 122.81 (CH-6); 125.88(CH-2-thienyl); 130.18 (CH-5-thienyl); 131.28 (CH-4-thienyl); 136.31(C-3-thienyl); 153.09 (C-7a); 154.20 (CH-2); 160.06 (C-4). ³¹P (′H dec.)NMR (202.4 MHz, D₂O+phosphate buffer, pH=7.1, ref_(H3PO4)=0 ppm): −21.14(bdd, J=21.0, 19.3, P_(β)); −10.39 (d, J=19.3, P_(α)); −6.45 (d, J=21.0,P_(γ)). MS (ESI, negative mode) m/z 609 (M−2Na+H), 631 (M−Na). HRMS(ESI, negative mode) for C₁₅H₁₇N₄NaO₁₃P₃S [M−2Na+H] calcd: 608.9618.found: 608.9635.

Example 354-Amino-5-phenyl-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-monophosphate sodium salt (14a)

Step 1. 4-Amino-5-iodo-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-monophosphate sodium salt (12)

Phosphorus oxychloride (70 μl, 0.77 mmol) was dropwise added to astirred mixture of 7-iodotubericidine 1 (250 mg, 0.64 mmol) in trimethylphosphate (2 ml) at 0° C. and the solution was stirred at 0° C. for 2 h.The reaction was quenched by the addition of aq TEAB (2M, 2 ml) andafter evaporation the rest was several times co-evaporated with water.The residue was purified by ion-exchange chromatography on DEAE-Sephadex(0→60% 2M aq TEAB in water) affording after ion-exchange on Dowex 50(Na⁺ form) and lyophilization title compound 12 as a white cotton (229mg, 73% yield). ¹H NMR (500 MHz, D₂O, ref_(dioxane)=3.75 ppm): 3.97 (dt,1H, J_(gem)=11.4, J_(H,P)=J_(5′b,4′)=4.0, H-5′b); 4.00 (ddd, 1H,J_(gem)=11.4,J_(H,P)=5.3, J_(5′a,4′)3.7, H-5′a); 4.30 (m, 1H,J_(4′,5′)=4.0, 3.7, J_(4′,3′)=3.0, J_(H,P)=1.0, H-4′); 4.44 (dd, 1H,J_(3′,2′)=5.4, =3.0, H-3′); 4.63 (dd, J_(2′,1′)=6.7, J_(2′,3′)=5.4,H-2′); 6.20 (d, 1H, J_(1′,2′)=6.7, H-1′); 7.70 (s, 1H, H-6); 8.08 (s,1H, H-2). ¹³C NMR (125.7 MHz, D₂O, ref_(dioxane)=69.3 ppm): 54.97 (C-5);66.81 (d, J_(C,P)=5, CH₂-5′); 73.50 (CH-3′); 76.59 (CH-2′); 86.74 (d,J_(C,P)=9, CH-4′); 88.52 (CH-1′); 106.71 (C-4a); 129.72 (CH-6); 152.45(C-7a); 154.24 (CH-2); 159.74 (C-4). ³¹P (¹H dec.) NMR (202.4 MHz, D₂O,ref_(H3PO4)=0 ppm): 3.24. MS (ESI, negative mode) m/z 471 (M−Na). HRMS(ESI, negative mode) for C₁₁H₁₃N₄IO₇P: [M−Na] calcd: 470.9561. found:470.9576.

Step 2.4-Amino-5-phenyl-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-monophosphate sodium salt (14a)

An argon purged mixture of Pd(OAc)₂ (1.7 mg, 7.6 μmol) and TPPTS (21.7mg, 38 μmol) in water/MeCN (2:1, 1.6 ml) was sonicated to dissolutionand quarter of resulting solution (0.4 ml, ¼ of total amount) was addedto an argon purged mixture of compound 12 from Step 1 (17 mg, 34 μmol),phenylboronic acid (7.9 mg, 65 μmol) and Na₂CO₃ (17 mg, 160 μmol) inwater/MeCN (2:1, 0.8 ml) and the mixture was stirred at 125° C. for 1.5h. After cooling, the mixture was filtered through microfilter andpurified by HPLC on C-18 phase (0→100% MeOH in 0.1M aq TEAB) affordingafter ion exchange on Dowex 50 (Na⁺ form) and lyophilization titlecompound 14a as white solid (14.4 mg, 94%). ¹H NMR (500 MHz, D₂O,ref_(dioxane)=3.75 ppm): 3.91 (dt, 1H, J_(gem)=11.4,J_(H,P)=J_(5′b,4′)4.4, H-5′b); 3.95 (ddd, 1H, J_(gem)=11.4, J_(H,P)=5.6,=4.2, H-5′a); 4.30 (ddd, 1H, J_(4′,5′)=4.4, 4.2, J_(4′,3′)=2.7, H-4);4.46 (dd, 1H, J_(3′,2′)=5.4, J_(3′,4′)=2.7, H-3); 4.75 (dd, 1H,J_(2′,1′)=7.1,J_(2′,3′)=5.4, H-2′); 6.31 (d, 1H, =7.1, H-1′); 7.46 (m,1H, H-p-Ph); 7.54 (m, 2H, H-m-Ph); 7.56 (m, 2H, H-o-Ph); 7.58 (s, 1H,H-6); 8.18 (s, 1H, H-2). ¹³C NMR (125.7 MHz, D₂O, ref_(dioxane)=69.3ppm): 66.53 (d, J_(C,P)=5, CH₂-5′); 73.61 (CH-3′); 76.15 (CH-2′); 86.94(d, J_(C,P)=9, CH-4′); 88.22 (CH-1); 103.97 (C-4a); 121.38 (C-5); 122.79(CH-6); 130.44 (CH-p-Ph); 131.72 (CH-o-Ph); 131.91 (CH-m-Ph); 136.31(C-i-Ph); 153.35 (C-7a); 154.40 (CH-2); 160.15 (C-4). ³¹P (¹H dec.) NMR(202.4 MHz, D₂O, ref_(H3PO4)=0 ppm): 4.64. MS (ESI) m/z 445 (M+H), 467(M+Na). HRMS (ESI) for C₁₇H₁₉N₄NaO₇P: [M+H] calcd: 445.0884. found:445.0880.

Example 364-Amino-5-(4-fluorophenyl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-monophosphate sodium salt (14b)

Title compound was prepared by following the procedure in Example 35,Step 2. White solid. Yield 47%. ¹H NMR (500 MHz, D₂O, ref_(dioxane)=3.75ppm): 3.90 (dt, 1H, J_(gem)=11.4, J_(H,P)=J_(5′b,4′)=4.1, H-5′b); 3.94(ddd, 1H, J_(gem)=11.4, J_(H,P)=5.6, J_(5′a,4′)=4.1, H-5′a); 4.30 (tdd,1H, J_(4′,5′)=4.1,J_(2′,3′)=2.7, J_(H,P)=1.1, H-4′); 4.46 (dd, 1H,J_(3′,2′)=5.4, J_(3′,4′)=2.7, H-3′); 4.74 (dd, 1H, J_(2′,1′)=7.2,J_(2′,3′)=5.4, H-2′); 6.30 (d, 1H, J_(1′,2′)=7.2, H-1′); 7.25 (m, 2H,H-m-C₆H₄F); 7.54 (m, 2H, H-o-C₆H₄F); 7.57 (s, 1H, H-6); 8.18 (s, 1H,H-2). ¹³C NMR (125.7 MHz, D₂O, ref_(dioxane)=69.3 ppm): 66.52 (d,J_(C,P)=4, CH₂-5′); 73.61 (CH-3′); 76.17 (CH-2′); 86.97 (d, J_(C,P)=9,CH-4′); 88.20 (CH-1′); 104.07 (C-4a); 118.55 (d, J_(C,F)=22,CH-m-C₆H₄F); 120.44 (C-5); 122.89 (CH-6); 132.35 (d, J_(C,F)=3,C-i-C₆H₄F); 133.55 (d, J_(C,F)=8, CH-o-C₆H₄F); 153.31 (C-7a); 154.45(CH-2); 160.18 (C-4); 165.00 (d, J_(C,F)=245, C-p-C₆H₄F). ³¹P (¹H dec.)NMR (202.4 MHz, D₂O, ref_(H3PO4)=0 ppm): 4.65. MS (ESI, negative mode)m/z 439 (M−Na), 461 (M−H). HRMS (ESI, negative mode) for C₁₇H₁₇FN₄O₇P[M−Na] calcd: 439.0813. found: 439.0828; for C₁₇H₁₆FN₄NaO₇P [M−H] calcd:461.0633. found: 461.0647.

Example 374-Amino-5-(furan-2-yl)-7-(6-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-monophosphate sodium salt (14c)

Title compound was prepared by following the procedure in Example 35,Step 2. Tan cotton. Yield 34%. ¹H NMR (500 MHz, D₂O, ref_(dioxane)=3.75ppm): 3.94 (ddd, 1H, J_(gem)=11.4, J_(H,P)=4.0, =2.6, H-5′b); 3.98 (ddd,1H, J_(gem)=11.4, J_(H,P)=5.2, J_(5′a,4′)=3.7, H-5′a); 4.31 (m, 1H,J_(4′,5′)=3.7, 2.6, J_(4′,3′)=2.9, J_(H,P)=1.1, H-4′); 4.47 (dd, 1H,J_(3′,2′)=5.3,J_(3′,4′)=2.9, H-3′); 4.73 (dd, 1H, J_(2′,1′)=6.9,J_(2′,3′)=5.3, H-2′); 6.28 (d, 1H, J_(1′,2′)=6.9, H-1′); 6.61 (dd, 1H,J_(4,3)=3.3, J_(4,5)=1.9, H-4-furyl); 6.78 (dd, 1H, J_(3,4)=3.3,J_(3,5)=0.7, H-3-furyl); 7.63 (dd, 1H, J_(5,4)=1.9, J_(5,3)=0.7,H-S-furyl); 7.87 (s, 1H, H-6); 8.14 (s, 1H, H-2). ¹³C NMR (125.7 MHz,D₂O, ref_(dioxane)=69.3 ppm): 66.52 (d, J_(C,P)=5, CH₂-5′); 73.65(CH-3′); 76.41 (CH-2′); 87.09 (d, J_(C,P)=9, CH-4′); 88.38 (CH-1′);102.81 (C-4a); 109.23 (CH-3-furyl); 111.01 (C-5); 114.78 (CH-4-furyl);122.30 (CH-6); 144.85 (CH-5-furyl); 150.56 (C-2-furyl); 153.34 (C-7a);154.57 (CH-2); 160.08 (C-4). ³¹P (¹H dec.) NMR (202.4 MHz, D₂O,ref_(H3PO4)=0 ppm): 4.37. MS (ESI, negative mode) m/z 411 (M−Na), 433(M−H). HRMS (ESI, negative mode) for C₁₅H₁₆N₄O₈P [M−Na] calcd: 411.0700.found: 411.0702.

Example 384-Amino-7-(8-D-ribofuranosyl)-5-(thiophen-2-yl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-monophosphate sodium salt (14d)

Title compound was prepared by following the procedure in Example 35,Step 2. Tan cotton. Yield 51%. ¹H NMR (500 MHz, D₂O, ref_(dioxane)=3.75ppm): 3.91 (dt, 1H, J_(gem)=11.4, J_(H,P)=J_(5′b,4′)=3.3, H-5′b); 3.95(ddd, 1H, J_(gem)=11.4, J_(H,P)=5.4, J_(5′a,4′)=4.2, H-5′a); 4.30 (ddd,1H, J_(4′,5′)=4.2, 3.3, J_(4′,3′)=2.8, H-4′); 4.45 (dd, 1H, =5.4,J_(3′,4′)=2.8, H-3); 4.72 (dd, 1H, J_(2′,1′)′=7.0, J_(2′,3′)=5.4, H-2′);6.28 (d, 1H, J_(3′,2′)=7.0, H-1′); 7.21 (dd, 1H, J_(4,5)=4.8,J_(4,3)=3.5, H-4-thienyl); 7.22 (dd, 1H, J_(3,4)=3.5, J_(3,5)=1.5,H-3-thienyl); 7.51 (dd, 1H, J_(5,4)=4.8, J_(5,3)=1.5, H-5-thienyl); 7.65(s, 1H, H-6); 8.17 (s, 1H, H-2). ¹³C NMR (125.7 MHz, D₂O,ref_(dioxane)=69.3 ppm): 66.53 (d, J_(C,P)=5, CH₂-5′); 73.57 (CH-3′);76.18 (CH-2′); 86.97 (d, J_(C,P)=9, CH-4′); 88.22 (CH-1′); 104.21(C-4a); 113.44 (C-5); 124.02 (CH-6); 129.30 (CH-5-thienyl); 130.25(CH-3-thienyl); 130.93 (CH-4-thienyl); 137.08 (C-2-thienyl); 153.12(C-7a); 154.63 (CH-2); 160.11 (C-4). ³¹P (¹H dec.) NMR (202.4 MHz, D₂O,ref_(H3PO4)=0 ppm): 4.57. MS (ESI, negative mode) m/z 427 (M−Na), 449(M−H). HRMS (ESI, negative mode) for C₁₅H₁₆SN₄O₇P [M−Na] calcd:427.0472. found: 427.0473.

Example 394-Amino-5-(furan-3-yl)-7-(β-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-monophosphate sodium salt (14e)

Title compound was prepared by following the procedure in Example 35,Step 2. White cotton. Yield 45%. ¹H NMR (500 MHz, D₂O,ref_(dioxane)=3.75 ppm): 3.91 (dt, 1H, J_(gem)=11.3,J_(H,P)=J_(5′b,4′)=4.1, H-5′b); 3.95 (ddd, 1H, J_(gem)=11.3,J_(H,P)=5.2, J_(5′a,4′)=4.1, H-5′a); 4.30 (td, 1H, J_(4′,5′)=4.1, =2.7,H-4′); 4.46 (dd, 1H, J_(3′,2′)=5.6, =2.7, H-3′); 4.73 (dd, 1H,J_(2′,1′)=7.0, J_(2′,3′)=5.6, H-2′); 6.28 (d, 1H, J_(1′,2′)=7.0, H-1′);6.73 (dd, 1H, J_(4,5)=1.8, J_(4,2)=0.9, H-4-furyl); 7.60 (s, 1H, H-6);7.66 (dd, 1H, J_(5,4)=1.8, J_(5,2)=1.6, H-5-furyl); 7.75 (dd, 1H,J_(2,5)=1.6, J_(2,4)=0.9, H-2-furyl); 8.17 (s, 1H, H-2). ¹³C NMR (151MHz, D₂O, ref_(dioxane)=69.3 ppm): 66.51 (d, J_(CP)=4, CH₂-5′); 73.66(CH-3′); 76.19 (CH-2′); 87.04 (d, J_(C,P)=9, CH-4′); 88.14 (CH-1′);104.48 (C-4a); 111.30 (C-5); 114.25 (CH-4-furyl); 120.37 (C-3-furyl);122.96 (CH-6); 143.22 (CH-2-furyl); 147.02 (CH-5-furyl); 153.24 (C-7a);154.46 (CH-2); 160.30 (C-4). ³¹P (¹H dec.) NMR (202.4 MHz, D₂O,ref_(H3PO4)=0 ppm): 4.55. MS (ESI, negative mode) m/z 411 (M−Na), 433(M−H). HRMS (ESI, negative mode) for C₁₅H₁₆N₄O₈P [M−Na] calcd: 411.0700.found: 411.0706; for C₁₅H₁₅N₄I₈PNa [M−H] calcd: 433.0520. found:411.0526.

Example 404-Amino-7-(β-D-ribofuranosyl)-5-(thiophen-3-yl)-7H-pyrrolo[2,3-d]pyrimidine5′-O-monophosphate sodium salt (14f)

Title compound was prepared by following the procedure in Example 35,Step 2. Pink solid. Yield 47%. ¹H NMR (600 MHz, D₂O, ref_(dioxane)=3.75ppm): 3.91 (dt, 1H, J_(gem)=11.4, J_(H,P)==4.1, H-5′b); 3.95 (ddd, 1H,J_(gem)=11.4, J_(H,P)=5.5, J_(5′a,4′)=4.0, H-5′a); 4.30 (m, 1H,J_(4′,5′)=4.1, 4.0, J_(4′,3′)=2.7, J_(H,P)=1.2, H-4′); 4.46 (dd, 1H,J_(3′,2′)=5.4,J_(3′,4′)=2.7, H-3); 4.74 (dd, 1H, J_(2′,1′)=7.1,J_(2′,3′)=5.4, H-2′); 6.29 (d, 1H, J_(1′,2′)=7.1, H-1); 7.32 (dd, 1H,J_(4,5)=4.9,J_(4,2)=1.4, H-4-thienyl); 7.51 (dd, 1H, J_(2,5)=2.9,J_(2,4)=1.4, H-2-thienyl); 7.608 (dd, 1H, J_(5,4)=4.9, J_(5,2)=2.9,H-5-thienyl); 7.612 (s, 1H, H-6); 8.16 (s, 1H, H-2). ¹³C NMR (151 MHz,D₂O, ref_(dioxane)=69.3 ppm): 66.55 (d, J_(C,P)=5, CH₂-5′); 73.66(CH-3′); 76.21 (CH-2′); 87.00 (d, J_(C,P)=9, CH-4′); 88.18 (CH-1′);104.18 (C-4a); 115.99 (C-5); 122.83 (CH-6); 125.98 (CH-2-thienyl);130.186 (CH-5-thienyl); 131.33 (CH-4-thienyl); 136.34 (C-3-thienyl);153.11 (C-7a); 154.39 (CH-2); 160.19 (C-4). ³¹P (¹H dec.) NMR (202.4MHz, D₂O, ref_(H3PO4)=0 ppm): 4.46. MS (ESI, negative mode) m/z 427(M−Na), 449 (M−H). HRMS (ESI, negative mode) for C₁₅H₁₆SN₄O₇P: [M−Na]calcd: 427.0472. found: 427.0474; for C₁₅H₁₅SN₄O₇PNa: [M−H] calcd:449.0291. found: 449.0295.

The invention has been described with reference to various specific andpreferred embodiments and techniques. However, it should be understoodthat many variations and modifications may be made while remainingwithin the spirit and scope of the invention.

What is claimed is:
 1. A compound of formula 1:

wherein: R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ is aryl,optionally substituted by one or two substituents selected from thegroup consisting of alkoxy, alkylthio, or halogen; R₃ is hydrogen oralkyl; or a pharmaceutically acceptable salt thereof; or an opticalisomer thereof; or a mixture of optical isomers.
 2. The compound ofclaim 1, wherein R₁ is hydrogen, mono-, di-, or tri-phosphate; R₂ isaryl that is optionally substituted by one substituent selected from thegroup consisting of alkoxy, alkylthio, or halogen; R₃ is hydrogen; or apharmaceutically acceptable salt thereof; or an optical isomer thereof;or a mixture of optical isomers.
 3. The compound of claim 2, wherein R₁is hydrogen; R₂ is phenyl that is optionally substituted by onesubstituent selected from the group consisting of (C₁-C₄) alkoxy,(C₁-C₄) alkylthio, or halogen; R₃ is hydrogen; or a pharmaceuticallyacceptable salt thereof; or an optical isomer thereof; or a mixture ofoptical isomers.
 4. The compound of claim 1, wherein R₁ is hydrogen,mono-, di-, or tri-phosphate; R₂ is aryl that is optionally substitutedby one substituent selected from the group consisting of alkoxy,alkylthio, or halogen; R₃ is alkyl; or a pharmaceutically acceptablesalt thereof; or an optical isomer thereof; or a mixture of opticalisomers.
 5. The compound of claim 4, wherein R₁ is hydrogen; R₂ isphenyl that is optionally substituted by one substituent selected fromthe group consisting of (C₁-C₄) alkoxy, (C₁-C₄) alkylthio, or halogen;R₃ is (C₁-C₄) alkyl; or a pharmaceutically acceptable salt thereof; oran optical isomer thereof; or a mixture of optical isomers.
 6. Thecompound of claim 1 being selected from the following compounds:

or a pharmaceutically acceptable salt thereof; or an optical isomerthereof; or a mixture of optical isomers.
 7. A method of inhibitingtumor/cancer growth in a subject comprising administering to the subjecta therapeutically effective amount of the compound according to claim 1.8. A method of inhibiting cell proliferation in tumor/cancer cells in asubject comprising administering to the subject a therapeuticallyeffective amount of the compound according to claim
 1. 9. A method oftreating a cellular proliferation disease in a subject comprisingadministering to the subject a therapeutically effective amount of thecompound according to claim
 1. 10. A method of treating a neoplasticdisease in a subject comprising administering to the subject atherapeutically effective amount of the compound according to claim 1.11. A method of treating a tumor or cancer in a subject comprisingadministering to the subject a therapeutically effective amount of thecompound according to claim
 1. 12. A pharmaceutical compositioncomprising a therapeutically effective amount of the compound accordingto claim 1 and one or more pharmaceutically acceptable carriers.
 13. Apharmaceutical composition comprising a therapeutically effective amountof the compound according to claim 1, and a second therapeutic agentselected from the group consisting of anthracyclines, DNA intercalators,alkylating agents, hormonal agents, LHRH agonists and antagonists,aromatase inhibitors, antiandrogens, chemoprevention agents, cell-cyclechemopreventative agents, antineoplasts, antimitotic agents, plantalkaloids, topoisomerase I inhibitors, topoisomerase II inhibitors,proteosomes inhibitors, nucleoside analogues, cytokines, growth factors,anti-angiogenic factors, and one or more pharmaceutically acceptablecarriers.